Azaindazole compounds and methods of use

ABSTRACT

Provided are methods for the treatment of rheumatoid arthritis using compounds of formula Ia and Ib: 
                         
or a pharmaceutically acceptable salt, hydrate or N-oxide thereof.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.11/752,201, filed May 22, 2007 now U.S. Pat. No. 7,777,035, which is acontinuation-in-part application of U.S. application Ser. No.11/474,132, filed Jun. 22, 2006 now U.S. Pat. No. 7,524,845 which claimspriority to U.S. Provisional Application No. 60/693,525 filed on Jun.22, 2005, the contents of each are incorporated herein in their entiretyfor all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

NOT APPLICABLE

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

NOT APPLICABLE

BACKGROUND OF THE INVENTION

The present invention provides compounds, pharmaceutical compositionscontaining one or more of those compounds or their pharmaceuticallyacceptable salts, which are effective in inhibiting the binding ofvarious chemokines, such as MIP-1α, leukotactin, MPIF-1 and RANTES, tothe CCR1 receptor. As antagonists or modulators for the CCR1 receptor,the compounds and compositions have utility in treating inflammatory andimmune disorder conditions and diseases.

Human health depends on the body's ability to detect and destroy foreignpathogens that might otherwise take valuable resources from theindividual and/or induce illness. The immune system, which comprisesleukocytes (white blood cells (WBCs): T and B lymphocytes, monocytes,macrophages granulocytes, NK cell, mast cells, dendritic cell, andimmune derived cells (for example, osteoclasts)), lymphoid tissues andlymphoid vessels, is the body's defense system. To combat infection,white blood cells circulate throughout the body to detect pathogens.Once a pathogen is detected, innate immune cells and cytotoxic T cellsin particular are recruited to the infection site to destroy thepathogen. Chemokines act as molecular beacons for the recruitment andactivation of immune cells, such as lymphocytes, monocytes andgranulocytes, identifying sites where pathogens exist.

Despite the immune system's regulation of pathogens, certaininappropriate chemokine signaling can develop and has been attributed totriggering or sustaining inflammatory disorders, such as rheumatoidarthritis, multiple sclerosis and others. For example, in rheumatoidarthritis, unregulated chemokine accumulation in bone joints attractsand activates infiltrating macrophages and T-cells. The activities ofthese cells induce synovial cell proliferation that leads, at least inpart, to inflammation and eventual bone and cartilage loss (see,DeVries, M. E., et al., Semin Immunol 11(2):95-104 (1999)). A hallmarkof some demyelinating diseases such as multiple sclerosis is thechemokine-mediated monocyte/macrophage and T cell recruitment to thecentral nervous system (see, Kennedy, et al., J. Clin Immunol.19(5):273-279 (1999)). Chemokine recruitment of destructive WBCs totransplants has been implicated in their subsequent rejection. See,DeVries, M. E., et al., ibid. Because chemokines play pivotal roles ininflammation and lymphocyte development, the ability to specificallymanipulate their activity has enormous impact on ameliorating andhalting diseases that currently have no satisfactory treatment. Inaddition, transplant rejection may be minimized without the generalizedand complicating effects of costly immunosuppressive pharmaceuticals.

Chemokines, a group of greater than 40 small peptides (7-10 kD), ligatereceptors expressed primarily on WBCs or immune derived cells, andsignal through G-protein-coupled signaling cascades to mediate theirchemoattractant and chemostimulant functions. Receptors may bind morethan one ligand; for example, the receptor CCR1 ligates RANTES(regulated on activation normal T cell expressed), MIP-1α (macrophageinflammatory protein), MPIF-1/CKβ8, and Leukotactin chemokines (amongothers with lesser affinities). To date, 24 chemokine receptors areknown. The sheer number of chemokines, multiple ligand bindingreceptors, and different receptor profiles on immune cells allow fortightly controlled and specific immune responses. See, Rossi, et al.,Ann. Rev. Immunol. 18(1):217-242 (2000). Chemokine activity can becontrolled through the modulation of their corresponding receptors,treating related inflammatory and immunological diseases and enablingorgan and tissue transplants.

The receptor CCR1 and its chemokine ligands, including, for exampleMIP-1α, MPIF-1/CKβ8, leukotactin and RANTES, represent significanttherapeutic targets (see Saeki, et al., Current Pharmaceutical Design9:1201-1208 (2003)) since they have been implicated in rheumatoidarthritis, transplant rejection (see, DeVries, M. E., et al., ibid.),and multiple sclerosis (see, Fischer, et al., J Neuroimmunol. 110(1-2):195-208 (2000); Izikson, et al., J. Exp. Med. 192(7):1075-1080(2000); and Rottman, et al., Eur. J. Immunol. 30(8):2372-2377 (2000). Infact, function-blocking antibodies, modified chemokine receptor ligandsand small organic compounds have been discovered, some of which havebeen successfully demonstrated to prevent or treat somechemokine-mediated diseases (reviewed in Rossi, et al., ibid.). Notably,in an experimental model of rheumatoid arthritis, disease development isdiminished when a signaling-blocking, modified-RANTES ligand isadministered (see Plater-Zyberk, et al., Immunol Lett. 57 (1-3):117-120(1997)). While function-blocking antibody and small peptide therapiesare promising, they suffer from the perils of degradation, extremelyshort half-lives once administered, and prohibitive expense to developand manufacture, characteristic of most proteins. Small organiccompounds are preferable since they often have longer half lives invivo, require fewer doses to be effective, can often be administeredorally, and are consequently less expensive. Some organic antagonists ofCCR1 have been previously described (see, Hesselgesser, et al., J. Biol.Chem. 273(25):15687-15692 (1998); Ng, et al., J. Med. Chem.42(22):4680-4694 (1999); Liang, et al., J. Biol. Chem.275(25):19000-19008 (2000); and Liang, et al., Eur. J. Pharmacol.389(1):41-49 (2000)). In view of the effectiveness demonstrated fortreatment of disease in animal models (see, Liang, et al., J. Biol.Chem. 275(25):19000-19008 (2000)), the search has continued to identifyadditional compounds that can be used in the treatment of diseasesmediated by CCR1 signaling.

BRIEF SUMMARY OF THE INVENTION

The present invention provides compounds having a formula selected fromthe group consisting of:

or a pharmaceutically acceptable salt, hydrate or N-oxide thereof. Inthe formulae above, the subscript m is an integer of from 0 to 4.

The symbol R¹ is a substituent independently selected from the groupconsisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, —CO₂R^(a),—S(O)₃R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a), —X¹S(O)₃R^(a), —X¹OR^(a),—COR^(a), —CONR^(a)R^(b), —X¹NR^(a)R^(b), —X¹NR^(a)COR^(b),—X¹CONR^(a)R^(b), X¹S(O)₂NR^(a)R^(b), X¹S(O)₂R^(a), —OR^(a),—NR^(a)R^(b), —NR^(a)COR^(b), —CONR^(a)R^(b), —NR^(a)S(O)₂R^(b),—S(O)₂NR^(a)R^(b), —S(O)₂R^(a), —X¹COR^(a), X¹CONR^(a)R^(b), and—X¹NR^(a)S(O)₂R^(b), wherein X¹ is C₁₋₄ alkylene and each R^(a) andR^(b) is independently selected from the group consisting of hydrogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl, or optionally R^(a) andR^(b) when attached to the same nitrogen atom are combined to form a 3-to 7-membered ring having from 0-2 additional heteroatoms as ringmembers; and wherein the aliphatic portions of each of said R¹substituents is optionally substituted with from one to three membersselected from the group consisting of —OH, —OR^(m), —OC(O)NHR^(m),—OC(O)N(R^(m))₂, —SH, —SR^(m), —S(O)R^(m), —S(O)₂R^(m), —SO₂NH₂,—S(O)₂NHR^(m), —S(O)₂N(R^(m))₂, —NHS(O)₂R^(m), —NR^(m)S(O)₂R^(m),—C(O)NH₂, —C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m), —NHC(O)R^(m),—NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂, —NR^(m)C(O)NHR^(m),—NHC(═NH)NH₂, —NHC(═NR^(m))NH₂, —NR^(m)C(═NR^(m))N(R^(m))₂,—NR^(m)C(═NR^(m))NH(R^(m)), —NHC(═NR^(m))NH(R^(m)),—NHC(═NR^(m))N(R^(m))₂, —NHC(═NH)N(R^(m))₂, —NHC(═NH)NH(R^(m)),—C(═NH)NH₂, —C(═NR^(m))NH₂, —C(═NR^(m))N(R^(m))₂, —C(═NR^(m))NH(R^(m)),—NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂, —NHC(O)N(R^(m))₂, —CO₂H,—CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN, —NO₂, —NH₂, —NHR^(m),—N(R^(m))₂, —NR^(m)S(O)NH₂ and —NR^(m)S(O)₂NHR^(m), wherein each R^(m)is independently an unsubstituted C₁₋₆ alkyl.

The symbols R^(2a), R^(2c) and R^(2d) in formulae Ia and Ib are eachsubstituents independently selected from the group consisting ofhydrogen, halogen, cyano, aryl, heteroaryl, —NO₂, —CO₂R^(c),—CONR^(c)R^(d), —C(O)R^(c), —S(O)R^(e), —S(O)₂R^(e), —S(O)₃R^(c),—R^(e), —C(NOR^(c))R^(d), —C(NR^(c)V)═NV, —N(V)C(R^(c))═NV,—X²C(NOR^(c))R^(d), —X²C(NR^(c)V)═NV, —X²N(V)C(R^(c))═NV,—X²NR^(c)R^(d), —X²SR^(c), —X²CN, —X²NO₂, —X²CO₂R^(c), —X²CONR^(c)R^(d),—X²C(O)R^(c), —X²OC(O)NR^(c)R^(d), —X²NR^(d)C(O)R^(c),—X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d), —X²NH—C(NH₂)═NH,—X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e), —X²NH—C(NHR^(e))═NH,—X²S(O)R^(e), —X²S(O)₂R^(e), —X²NR^(c)S(O)₂R^(e), —X²S(O)₂NR^(c)R^(d),—X²N₃, —SR^(c), —NR^(d)C(O)R^(c), —NR^(d)C(O)₂R^(c), —X²S(O)₃R^(c),—S(O)₂NR^(c)R^(d), —X²OR^(c), —O—X²OR^(c), —X²NR^(c)R^(d),—O—X²NR^(c)R^(d), —NR^(d)—X²CO₂R^(c), —NR^(c)—C(O)NR^(c)R^(d),—NH—C(NH₂)═NH, —NR^(e)C(NH₂)═NH, —NH—C(NH₂)═NR^(e), —NH—C(NHR^(e))═NH,—NR^(e)C(NHR^(e))═NH, —NR^(e)C(NH₂)═NR^(e), —NH—C(NHR^(e))═NR^(e),—NH—C(NR^(e)R^(e))═NH, NR^(c)S(O)₂R^(e), —NR^(c)C(S)NR^(c)R^(d),—X²NR^(c)C(S)NR^(c)R^(d), —X²OC(O)R^(c), —O—X²CONR^(c)R^(d),—OC(O)R^(c), —NR^(c)R^(d), —NR^(d)—X²OR^(c) and —NR^(d)—X²NR^(c)R^(d).

Within each of R^(2a), R^(2c) and R^(2d), X² is C₁₋₄ alkylene and eachR^(c) and R^(d) is independently selected from hydrogen, C₁₋₈ alkyl,C₁₋₈ haloalkyl, and C₃₋₆ cycloalkyl. Optionally, R^(c) and R^(d) whenattached to the same nitrogen atom can be combined with the nitrogenatom to form a five or six-membered ring having from 0 to 2 additionalheteroatoms as ring members. The symbol R^(e) is independently selectedfrom the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl and heteroaryl, and each ofR^(c), R^(d) and R^(e) is optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(n),—OC(O)NHR^(n), —OC(O)N(R^(n))₂, —SH, —S(O)R^(n), —S(O)₂R^(n), —SO₂NH₂,—S(O)₂NHR^(n), —S(O)₂N(R^(n))₂, —NHS(O)₂R^(n), —NR^(n)S(O)₂R^(n),—C(O)NH₂, —C(O)NHR^(n), —C(O)N(R^(n))₂, —C(O)R^(n), —NHC(O)R^(n),—NR^(n)C(O)R^(n), —NHC(O)NH₂, —NR^(n)C(O)NH₂, —NR^(n)C(O)NHR^(n),—NHC(O)NHR^(n), —NR^(n)C(O)N(R^(n))₂, —NHC(O)N(R^(n))₂, —CO₂H,—CO₂R^(n), —NHCO₂R^(n), —NR^(n)CO₂R^(n), —CN, —NO₂, —NH₂, —NHR^(n),—N(R^(n))₂, —NR^(n)S(O)NH₂ and —NR^(n)S(O)₂NHR^(n), wherein each R^(n)is independently an unsubstituted C₁₋₆ alkyl; and wherein V isindependently selected from the group consisting of —R^(c), —CN,—CO₂R^(e) and —NO₂.

Each of ring vertices a, b, c and d in formulae Ia and Ib isindependently selected from N and C(R^(3a)), and from one to two of saidring vertices is N. The symbol R^(3a) in formulae Ia and Ib isindependently selected from the group consisting of hydrogen, halogen,—OR^(f), —OC(O)R^(f), —NR^(f)R^(g), —SR^(f), —R^(h), —CN, —NO₂,—CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f), —OC(O)NR^(f)R^(g),—NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h), —NR^(f)—C(O)NR^(f)R^(g),—NH—C(NH₂)═NH, —NR^(h)C(NH₂)═NH, —NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH,—C(═NR^(f))NR^(g)R^(h), —S(O)₃R^(f), —S(O)R^(h), —S(O)₂R^(h),—S(O)₃R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —NR^(f)S(O)₂R^(h),—NR^(f)S(O)₂NR^(f)R^(g), —N₃, —C(C═NOR^(f))NR^(f)R^(g), —X₃SO₃R^(f),—X³C(═NR^(f))NR^(g)R^(h), —X³OR^(f), —X³OC(O)R^(f), —X³NR^(f)R^(g),—X³SR^(f), —X³CN, —X³NO₂, —X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f),—X³OC(O)NR^(f)R^(g), —X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h),—X³NR^(f)—C(O)NR^(f)R^(g), —X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH,—X³NH—C(NH₂)═NR^(h), —X³NH—C(NHR^(h))═NH, —X³S(O)R^(h), —X³S(O)₂R^(h),—X³NR^(f)S(O)₂R^(h), —X³S(O)₂NR^(f)R^(g), —Y, —X³Y, —X³N₃,—C(O)NR^(f)S(O)R^(h), —P═O(OR^(f))(OR^(g)), —X³C(O)NR^(f)S(O)₂R^(h),—X³C(O)NR^(f)S(O)R^(h) and —X³P═O(OR^(f))(OR^(g)). The symbol Y is afive to ten-membered aryl, heteroaryl or heterocycloalkyl ring,optionally substituted with from one to three substitutents selectedfrom the group consisting of halogen, —OR^(f), —NR^(f)R^(g), —R^(h),—SR^(f), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h),—S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g), —X³NR^(f)S(O)₂R^(h) and—X³S(O)₂NR^(f)R^(g), and wherein each X³ is independently selected fromthe group consisting of C₁₋₄ alkylene, C₂₋₄ alkenylene and C₂₋₄alkynylene; each R^(f) and R^(g) is independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄ alkyl, and aryloxy-C₁₋₄ alkyl,or when attached to the same nitrogen atom can be combined with thenitrogen atom to form a five or six-membered ring having from 0 to 2additional heteroatoms as ring members; and each R^(h) is independentlyselected from the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄alkyl, and aryloxy-C₁₋₄ alkyl, wherein the aliphatic portions of X³,R^(f), R^(g) and R^(h) are optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R)₂, —NHS(O)₂R^(o), —NR^(o)S(O)₂R^(o),—C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o), —NHC(O)R^(o),—NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂, —NR^(o)C(O)NHR^(o),—NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂, —NHC(O)N(R^(o))₂, —CO₂H,—CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN, —NO₂, —NH₂, —NHR^(o),—N(R^(o))₂, —NR^(o)S(O)NH₂ and —NR^(o)S(O)₂NHR^(o), wherein R^(o) isunsubstituted C₁₋₆ alkyl.

In addition to the compounds provided herein, the present inventionfurther provides pharmaceutical compositions containing one or more ofthese compounds, as well as methods for the use of these compounds intherapeutic methods, primarily to treat diseases associated with CCR1signaling activity.

BRIEF DESCRIPTION OF THE DRAWINGS

NONE

DETAILED DESCRIPTION OF THE INVENTION I. Abbreviation and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkyl” refers to a cycloalkyl group that contain from one tofive heteroatoms selected from N, O, and S, wherein the nitrogen andsulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. The heterocycloalkyl may be a monocyclic, abicyclic or a polycyclic ring system. Non limiting examples ofheterocycloalkyl groups include pyrrolidine, piperidinyl, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrahydrothiophene, quinuclidine, and the like. Aheterocycloalkyl group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as —NR^(a)R^(b) ismeant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl andthe like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like).

The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in someembodiments, will include both substituted and unsubstituted forms ofthe indicated radical. Preferred substituents for each type of radicalare provided below. For brevity, the terms aryl and heteroaryl willrefer to substituted or unsubstituted versions as provided below, whilethe term “alkyl” and related aliphatic radicals is meant to refer tounsubstituted version, unless indicated to be substituted.

Substituents for the alkyl radicals (including those groups oftenreferred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: -halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2 m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedheteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens,unsubstituted C₁₋₈ alkyl, C₁₋₈ alkoxy or C₁₋₈ thioalkoxy groups, orunsubstituted aryl-C₁₋₄ alkyl groups. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to fauna 3-, 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant toinclude 1-pyrrolidinyl and 4-morpholinyl.

Similarly, substituents for the aryl and heteroaryl groups are variedand are generally selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′,—R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C₁₋₄ alkyl, andunsubstituted aryloxy-C₁₋₄ alkyl. Other suitable substituents includeeach of the above aryl substituents attached to a ring atom by analkylene tether of from 1-4 carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. The compounds of the present invention may alsocontain unnatural proportions of atomic isotopes at one or more of theatoms that constitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

II. General

The present invention derives from the discovery that compounds offormula Ia or Ib (as well as the subgeneric formulae Ia¹⁻⁴ and Ib¹⁻⁴)act as potent antagonists of the CCR1 receptor. The compounds have invivo anti-inflammatory activity. Accordingly, the compounds providedherein are useful in pharmaceutical compositions, methods for thetreatment of CCR1-mediated diseases, and as controls in assays for theidentification of competitive CCR1 antagonists.

III. Compounds

In one aspect, the present invention provides compounds having a formulaselected from the group consisting of:

or a pharmaceutically acceptable salt, hydrate or N-oxide thereof. Inthe formulae above, the subscript m is an integer of from 0 to 4. Incertain embodiments, in formulae Ia and Ib the subscript m is an integerfrom 0 to 2. In yet another embodiment, the subscript m in formulae Iaand Ib is an integer of from 0 to 1.

The symbol R¹ in formulae Ia and Ib is a substituent independentlyselected from the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, —CO₂R^(a), —S(O)₃R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a),—X¹S(O)₃R^(a), —X¹OR^(a), —COR^(a), —CONR^(a)R^(b), —X¹NR^(a)R^(b),—X¹NR^(a)COR^(b), —X¹CONR^(a)R^(b), X¹S(O)₂NR^(a)R^(b), X¹S(O)₂R^(a),—OR^(a), —NR^(a)R^(b), —NR^(a)COR^(b), —CONR^(a)R^(b),—NR^(a)S(O)₂R^(b), —S(O)₂NR^(a)R^(b), —S(O)₂R^(a), —X¹COR^(a),X¹CONR^(a)R^(b), and —X¹NR^(a)S(O)₂R^(b). The symbol X¹ is C₁₋₄ alkyleneand each R^(a) and R^(b) substituent is independently selected from thegroup consisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl, or optionally R^(a) and R^(b) when attached to the samenitrogen atom are combined to form a 3- to 7-membered ring having from0-2 additional heteroatoms as ring members; and wherein the aliphaticportions of each of said R¹ substituents is optionally substituted withfrom one to three members selected from the group consisting of —OH,—OR^(m), —OC(O)NHR^(m), —OC(O)N(R^(m))₂, —SH, —SR^(m), —S(O)R^(m),—S(O)₂R^(m), —SO₂NH₂, —S(O)₂NHR^(m), —S(O)₂N(R^(m))₂, —NHS(O)₂R^(m),—NR^(m)S(O)₂R^(m), —C(O)NH₂, —C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m),—NHC(O)R^(m), —NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂,—NR^(m)C(O)NHR^(m), —NHC(═NH)NH₂, —NHC(═NR^(m))NH₂,—NR^(m)C(═NR^(m))N(R^(m))₂, —NR^(m)C(═NR^(m))NH(R^(m)),—NHC(═NR^(m))NH(R^(m)), —NHC(═NR^(m))N(R^(m))₂, —NHC(═NH)N(R^(m))₂,—NHC(═NH)NH(R^(m)), —C(═NH)NH₂, —C(═NR^(m))NH₂, —C(═NR^(m))N(R^(m))₂,—C(═NR^(m))NH(R^(m)), —NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂,—NHC(O)N(R^(m))₂, —CO₂H, —CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN,—NO₂, —NH₂, —NHR^(m), —N(R^(m))₂, —NR^(m)S(O)NH₂ and—NR^(m)S(O)₂NHR^(m), wherein each R^(m) is independently anunsubstituted C₁₋₆ alkyl.

In another embodiment, R¹ in formulae Ia and Ib is a substituentindependently selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl, C₃₋₆ cycloalkyl, —CO₂R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a) and—X¹OR^(a), wherein the aliphatic portions of each of said R¹substituents is optionally substituted with from one to three membersselected from the group consisting of —OH, —OR^(m), —OC(O)NHR^(m),—OC(O)N(R^(m))₂, —SH, —SR^(m), —S(O)R^(m), —S(O)₂R^(m), —SO₂NH₂,—S(O)₂NHR^(m), —S(O)₂N(R^(m))₂, —NHS(O)₂R^(m), —NR^(m)S(O)₂R^(m),—C(O)NH₂, —C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m), —NHC(O)R^(m),—NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂, —NR^(m)C(O)NHR^(m),—NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂, —NHC(O)N(R^(m))₂, —CO₂H,—CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN, —NO₂, —NH₂, —NHR^(m),—N(R^(m))₂, —NR^(m)S(O)NH₂ and —NR^(m)S(O)₂NHR^(m), wherein each R^(m)is independently an unsubstituted C₁₋₆ alkyl.

In another embodiment, R¹ in formulae Ia and Ib is a substituentindependently selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl and C₃₋₆ cycloalkyl, wherein the aliphatic portions of each ofsaid R¹ substituents is optionally substituted with from one to threemembers selected from the group consisting of —OH, —OR^(m),—OC(O)NHR^(m), —OC(O)N(R^(m))₂, —SH, —SR^(m), —S(O)R^(m), —S(O)₂R^(m),—SO₂NH₂, —S(O)₂NHR^(m), —S(O)₂N(R^(m))₂, —NHS(O)₂R^(m),—NR^(m)S(O)₂R^(m), —C(O)NH₂, —C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m),—NHC(O)R^(m), —NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂,—NR^(m)C(O)NHR^(m), —NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂,—NHC(O)N(R^(m))₂, —CO₂H, —CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN,—NO₂, —NH₂, —NHR^(m), —N(R^(m))₂, —NR^(m)S(O)NH₂ and—NR^(m)S(O)₂NHR^(m), wherein each R^(m) is independently anunsubstituted C₁₋₆ alkyl.

In one embodiment of the invention, R¹ in formulae Ia and Ib, ifpresent, is selected from the group consisting of —CO₂H or C₁₋₄ alkyl,optionally substituted with —OH, —OR^(m), —S(O)₂R^(m), —CO₂H and—CO₂R^(m). In another embodiment of the invention, R¹ is methyl; and mis 0-2.

The symbols R^(2a), R^(2c) and R^(2d) in formulae Ia and Ib are eachsubstituents independently selected from the group consisting ofhydrogen, halogen, cyano, aryl, heteroaryl, —NO₂, —CO₂R^(c),—CONR^(c)R^(d), —C(O)R^(c), —S(O)R^(e), —S(O)₂R^(e), —S(O)₃R^(c),—R^(e), —C(NOR^(c))R^(d), —C(NR^(c)V)═NV, —N(V)C(R^(c))═NV,—X²C(NOR^(c))R^(d), —X²C(NR^(c)V)═NV, —X²N(V)C(R^(c))═NV,—X²NR^(c)R^(d), —X²SR^(c), —X²CN, —X²NO₂, —X²CO₂R^(c), —X²CONR^(c)R^(d),—X²C(O)R^(c), —X²OC(O)NR^(c)R^(d), —X²NR^(d)C(O)R^(c),—X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d), —X²NH—C(NH₂)═NH,—X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e), —X²NH—C(NHR^(e))═NH,—X²S(O)R^(e), —X²S(O)₂R^(e), —X²NR^(c)S(O)₂R^(e), —X²S(O)₂NR^(c)R^(d),—X²N₃, —OR^(c), —SR^(c), —NR^(d)C(O)R^(c), —NR^(d)C(O)₂R^(e),—X²S(O)₃R^(c), —S(O)₂NR^(c)R^(d), —X²OR^(c), —O—X²OR^(c),—X²NR^(c)R^(d), —O—X²NR^(c)R^(d), —NR^(d)—X²CO₂R^(c),—NR^(c)—C(O)NR^(c)R^(d), —NH—C(NH₂)═NH, —NR^(e)C(NH₂)═NH,—NH—C(NH₂)═NR^(e), —NH—C(NHR^(e))═NH, —NR^(e)C(NHR^(e))═NH,—NR^(e)C(NH₂)═NR^(e), —NH—C(NHR^(e))═NR^(e), —NH—C(NR^(e)R^(e))═NH,NR^(c)S(O)₂R^(e), —NR^(c)C(S)NR^(c)R^(d), —X²NR^(c)C(S)NR^(c)R^(d),—X²OC(O)R^(c), —O—X²CONR^(c)R^(d), —OC(O)R^(c), —NR^(c)R^(d),—NR^(d)—X²OR^(c) and —NR^(d)—X²NR^(c)R^(d).

In one embodiment, the symbol R^(2a) in formulae Ia and Ib isindependently selected from the group consisting of hydrogen, halogen,cyano, heteroaryl, —NO₂, —CO₂R^(c), —CONR^(c)R^(d), —C(O)R^(c),—S(O)R^(e), —S(O)₂R^(e), —R^(e), —C(NOR^(c))R^(d), —C(NR^(c)V)═NV,—N(V)C(R^(c))═NV, —X²C(NOR^(c))R^(d), —X²C(NR^(c)V)═NV, —X²N(V)C(R^(c))═NV, —X²NR^(c)R^(d), —X²SR^(c), —X²CN, —X²NO₂, —X²CO₂R^(c),—X²CONR^(c)R^(d), —X²C(O)R^(c), —X²OC(O)NR^(c)R^(d), —X²NR^(d)C(O)R^(c),—X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d), —X²NH—C(NH₂)═NH,—X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e), —X²NH—C(NHR^(e))═NH,—X²S(O)R^(e), —X²S(O)₂R^(e), —X²NR^(c)S(O)₂R^(e), —X²S(O)₂NR^(c)R^(d)and —X²N₃.

In another embodiment, the R^(2a) substitutent in formulae Ia and Ib isselected from the group consisting of hydrogen, F, Cl, Br, I, —CO₂R^(c),—CONR^(c)R^(d), —CN, a 5- to 6-membered heteroaryl, —X₂NR^(c)R^(d),—C(NOR^(c))R^(d). In yet another embodiment, R^(2a) is hydrogen. In yetanother embodiment, the R^(2a) substitutent in formulae Ia and Ib isselected from the group consisting of F, Cl, Br, I, —CO₂Me, —CONH₂, CN,oxazolyl, —CH₂NH₂, —CH₂NHMe, —CH₂NMe₂ and —CH═N—OH. In yet anotherembodiment, in compounds having formulae Ia and Ib, the R^(2a)substituent is selected from the group consisting of hydrogen, F, Cl, Brand I.

In another embodiment, the symbols R^(2c) and R^(2d) in formulae Ia andIb are each substituents independently selected from the groupconsisting of halogen, —OR^(c), —SR^(c), —OC(O)R^(c), —NR^(c)R^(d),—R^(e), —CN, —NO₂, —CO₂R^(c), —C(O)R^(c), —NR^(d)C(O)R^(c),—NR^(d)C(O)₂R^(e), —S(O)₂R^(e), —S(O)₂NR^(c)R^(d), —X²OR^(c),—O—X²OR^(c), —X²NR^(c)R^(d), —O—X²NR^(c)R^(d) and —NR^(d)—X²CO₂R^(c). Incertain aspects of this embodiment, R^(2c) and R^(2d) are eachindependently selected from the group consisting of hydrogen, halogen,F, Cl, Br, I and OR^(c).

Within each of R^(2a), R^(2c) and R^(2d), X² is C₁₋₄ alkylene and eachR^(c) and R^(d) is independently selected from hydrogen, C₁₋₈ alkyl,C₁₋₈ haloalkyl, and C₃₋₆ cycloalkyl. Optionally, R^(c) and R^(d) whenattached to the same nitrogen atom can be combined with the nitrogenatom to form a five or six-membered ring having from 0 to 2 additionalheteroatoms as ring members. The symbol R^(e) is independently selectedfrom the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl and heteroaryl, and each ofR^(c), R^(d) and R^(e) is optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(n),—OC(O)NHR^(n), —OC(O)N(R^(n))₂, —SH, —SR^(n), —S(O)R^(n), —S(O)₂R^(n),—SO₂NH₂, —S(O)₂NHR^(n), —S(O)₂N(R^(n))₂, —NHS(O)₂R^(n),—NR^(n)S(O)₂R^(n), —C(O)NH₂, —C(O)NHR^(n), —C(O)N(R^(n))₂, —C(O)R^(n),—NHC(O)R^(n), —NR^(n)C(O)R^(n), —NHC(O)NH₂, —NR^(n)C(O)NH₂,—NR^(n)C(O)NHR^(n), —NHC(O)NHR^(n), —NR^(n)C(O)N(R^(n))₂,—NHC(O)N(R^(n))₂, —CO₂H, —CO₂R^(n), —NHCO₂R^(n), —NR^(n)CO₂R^(n), —CN,—NO₂, —NH₂, —NHR^(n), —N(R^(n))₂, —NR^(n)S(O)NH₂ and—NR^(n)S(O)₂NHR^(n), wherein each R^(n) is independently anunsubstituted C₁₋₆ alkyl; and wherein V is independently selected fromthe group consisting of —R^(c), —CN, —CO₂R^(e) and —NO₂.

In a certain embodiment of a compound having formulae Ia and Ib, thesubscript m is 0 or 1; and the symbol R^(2a) is hydrogen. In anotherembodiment, the subscript m is 0-1; and R^(2a) is F or Cl.

In another embodiment of the invention, R^(2c) in formulae Ia and Ib isselected from the group consisting of halogen, —CN, —NO₂, —CO₂R^(c),—COR^(c), —S(O)2R^(e). In another embodiments of the invention, thesymbol R^(2c) is selected from the group consisting of F, Cl, Br, CN,NO₂, —CO₂CH₃, —C(O)CH₃ and —S(O)₂CH₃.

In yet another embodiment of the invention, the symbol R^(2d) informulae Ia and Ib is selected from the group consisting of —SR^(c),—O—X²—OR^(c), —X²—OR^(c), —OC(O)R^(c), —NR^(c)R^(d), —R^(e) and —OR^(c).In another embodiment, R^(2d) is selected from the group consisting of—SMe, —OCH₂OMe, —CH₂OMe, —CH₂OEt, methyl, ethyl, methoxy and ethoxy.

In formulae Ia and Ib, each of the ring vertices a, b, c and d isindependently selected from N and C(R^(3a)), and from one to two of saidring vertices is N. In one embodiment of the invention, the fused sixmembered ring having vertices a, b, c and d is a fused pyridine ring ora fused pyrimidine ring. In yet another embodiment of the invention, thefused six membered ring having vertices a, b, c and d is a fusedpyrazine ring. In yet another embodiment of the invention, the fused sixmembered ring having vertices a, b, c and d is a fused pyridazine ring.

Turning to the R^(3a) substituent in formulae Ia and Ib, at eachoccurrence, the symbol R^(3a) is independently selected from the groupconsisting of hydrogen, halogen, —OR^(f), —OC(O)R^(f), —NR^(f)R^(g),—SR^(f), —R^(h), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—OC(O)NR^(f)R^(g), —NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h),—NR^(f)—C(O)NR^(f)R^(g), —NH—C(NH₂)═NH, —NR^(h)C(NH₂)═NH,—NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH, —C(═NR^(f))NR^(g)R^(h),—S(O)₃R^(f), —S(O)R^(h), —S(O)₂R^(h), —S(O)₃R^(h), —NR^(f)S(O)₂R^(h),—S(O)₂NR^(f)R^(g), —NR^(f)S(O)₂R^(h), —NR^(f)S(O)₂NR^(f)R^(g), —N₃,—C(C═NOR^(f))NR^(f)R^(g), —X₃SO₃R^(f), —X³C(═NR^(f))NR^(g)R^(h),—X³OR^(f), —X³OC(O)R^(f), —X³NR^(f)R^(g), —X³SR^(f), —X³CN, —X³NO₂,—X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f), —X³OC(O)NR^(f)R^(g),—X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h), —X³NR^(f)—C(O)NR^(f)R^(g),—X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH, —X³NH—C(NH₂)═NR^(h),—X³NH—C(NHR^(h))═NH, —X³S(O)R^(h), —X³S(O)₂R^(h), —X³NR^(f)S(O)₂R^(h),—X³S(O)₂NR^(f)R^(g), —Y, —X³Y, —X³N₃, —C(O)NR^(f)S(O)R^(h),—P═O(OR^(f))(OR^(g)), —X³C(O)NR^(f)S(O)₂R^(h), —X³C(O)NR^(f)S(O)R^(h)and —X³P═O(OR^(f))(OR^(g)). The symbol Y is a five to ten-membered aryl,heteroaryl or heterocycloalkyl ring, optionally substituted with fromone to three substitutents selected from the group consisting ofhalogen, —OR^(f), —NR^(f)R^(g), —R^(h), —SR^(f), —CN, —NO₂, —CO₂R^(f),—CONR^(f)R^(g), —C(O)R^(f), —NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h),—NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g),—X³NR^(f)S(O)₂R^(h) and —X³S(O)₂NR^(f)R^(g), and wherein each X³ isindependently selected from the group consisting of C₁₋₄ alkylene, C₂₋₄alkenylene and C₂₋₄ alkynylene; each R^(f) and R^(g) is independentlyselected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl,C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄ alkyl, andaryloxy-C₁₋₄ alkyl, or when attached to the same nitrogen atom can becombined with the nitrogen atom to form a five or six-membered ringhaving from 0 to 2 additional heteroatoms as ring members; and eachR^(h) is independently selected from the group consisting of C₁₋₈ alkyl,C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl,heteroaryl, aryl-C₁₋₄ alkyl, and aryloxy-C₁₋₄ alkyl, wherein thealiphatic portions of X³, R^(f), R^(g) and R^(h) are optionally furthersubstituted with from one to three members selected from the groupconsisting of —OH, —OR^(o), —OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH,—SR^(o), —S(O)R^(o), —S(O)₂R^(o), —SO₂NH₂, —S(O)₂NHR^(o),—S(O)₂N(R^(o))₂, —NHS(O)₂R^(o), —NR^(o)S(O)₂R^(o), —C(O)NH₂,—C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o), —NHC(O)R^(o),—NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂, —NR^(o)C(O)NHR^(o),—NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂, —NHC(O)N(R^(o))₂, —CO₂H,—CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN, —NO₂, —NH₂, —NHR^(o),—N(R^(o))₂, —NR^(o)S(O)NH₂ and —NR^(o)S(O)₂NHR^(o), wherein R^(o) isunsubstituted C₁₋₆ alkyl.

In one embodiment of formulae Ia and Ib, the symbol R^(3a), at eachoccurrence, is independently selected from the group consisting ofhydrogen, halogen, —OR^(f), —OC(O)R^(f), —NR^(f)R^(g), —SR^(f), —R^(h),—CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f), —OC(O)NR^(f)R^(g),—NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h), —NR^(f)—C(O)NR^(f)R^(g),—S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g),—NR^(f)S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g), —X³SR^(f), —X³CN,—X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f), —X³NR^(g)C(O)R^(f),—X³NR^(g)C(O)₂R^(h), —Y, —X³Y and —X³N₃. The symbol Y is a five orsix-membered aryl, a five or six membered heteroaryl, or a three toeight membered heterocycloalkyl ring, optionally substituted with fromone to three substitutents selected from the group consisting ofhalogen, —OR^(f), —NR^(f)R^(g), —R^(h), —SR^(f), —CN, —NO₂, —CO₂R^(f),—CONR^(f)R^(g), —C(O)R^(f), —NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h),—NR^(f)S(O)₂R^(h) and —S(O)₂NR^(f)R^(g). X³ is independently C₁₋₄alkylene. The symbols R^(f) and R^(g) are independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl, and each R^(h)is independently selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl and C₃₋₆ cycloalkyl. The aliphatic portions of X³, R^(f),R^(g) and R^(h) is optionally further substituted with from one to threemembers selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein each R^(o) is independently anunsubstituted C₁₋₆ alkyl.

In another embodiment of the invention, the symbol R^(3a) of formulae Iaand Ib is a member independently selected from the group consisting ofhydrogen, halogen, —OR^(f), —NR^(f)R^(g), —R^(h), —CN, and —Y, wherein Yis a five to six-membered aryl ring, a five to six-membered heteroarylring, or a three to eight-membered heterocycloalkyl ring selected fromthe group consisting of homopiperidinyl, morpholinyl, thiomorpholinyl,pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl,piperazinyl, phenyl, pyridyl, pyrimidinyl, oxadiazolyl, oxazolyl andthiazolyl, optionally substituted with from one to three substitutentsselected from the group consisting of halogen, —OR^(f), —NR^(f)R^(g),—R^(h), —CN, wherein each R^(f) and R^(g) is independently selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl and C₃₋₆ cycloalkyl, and each R^(h)is independently selected from the group consisting of C₁₋₆ alkyl, C₁₋₆haloalkyl and C₃₋₆ cycloalkyl, wherein the aliphatic portions of R^(f),R^(g) and R^(h) are optionally further substituted with from one tothree members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein R^(o) is unsubstituted C₁₋₆ alkyl.

In another embodiment of the invention, the R^(3a) groups in formulae Iaand Ib is selected from the group consisting of —Y and —X³—Y, wherein Yis selected from the group consisting of homopiperidinyl, morpholinyl,thiomorpholinyl, pyrrolidinyl, piperidinyl, azetidinyl, pyranyl,tetrahydrofuranyl, piperazinyl, phenyl, thienyl, furanyl, pyridyl,pyrimidinyl, pyrazinyl, pyrrolyl, pyridizinyl, pyrazolyl, imidazolyl,thiazolyl, oxazolyl, isoxazolyl, isothiazolyl, triazolyl, tetrazolyl andoxadiazolyl, which is optionally substituted with from one to threesubstituents independently selected from the group consisting ofhalogen, —OR^(f), —NR^(f)R^(g), —COR^(f), —CO₂R^(f), —CONR^(f)R^(g),—NO₂, —R^(h) and —CN, wherein R^(f) and R^(g) are each independentlyselected from the group consisting of H, C₁₋₈ alkyl, C₃₋₆ cycloalkyl andC₁₋₈ haloalkyl, and each R^(h) is independently selected from the groupconsisting of C₁₋₈ alkyl, C₃₋₆ cycloalkyl and C₁₋₈ haloalkyl. In certainembodiments of the invention, the symbol Y is selected from the groupconsisting of imidazolyl, phenyl, pyridyl, oxazolyl, pyrimidinyl,oxadiazolyl, and thiazolyl, each of which is optionally substituted withfrom one to three substituents independently selected from the groupconsisting of halogen, —OR^(f), —NR^(f)R^(g), —COR^(f), —CO₂R^(f),—CONR^(f)R^(g), —NO₂, —R^(h) and —CN, wherein R^(f) and R^(g) are eachindependently selected from the group consisting of H, C₁₋₈ alkyl, C₃₋₆cycloalkyl and C₁₋₈ haloalkyl, and each R^(h) is independently selectedfrom the group consisting of C₁₋₈ alkyl, C₃₋₆ cycloalkyl and C₁₋₈haloalkyl. In certain other embodiments of the invention, the symbol Yis selected from the group consisting of phenyl, pyridyl, oxazolyl,pyrimidinyl, oxadiazolyl, and thiazolyl, each of which is optionallysubstituted with from one to three substituents independently selectedfrom the group consisting of halogen, —OR^(f), —NR^(f)R^(g), —COR^(f),—CO₂R^(f), —CONR^(f)R^(g), —NO₂, —R^(h) and —CN, wherein R^(f) and R^(g)are each independently selected from the group consisting of H, C₁₋₈alkyl, C₃₋₆ cycloalkyl and C₁₋₈ haloalkyl, and each R^(h) isindependently selected from the group consisting of C₁₋₈ alkyl, C₃₋₆cycloalkyl and C₁₋₈ haloalkyl. Within this embodiment, in certainaspects of the invention, m is an integer from 0-2. In other aspect, mis an integer from 0-1.

In yet another embodiment of the invention, the R^(3a) substituent informulae Ia and Ib is selected from the group consisting of hydrogen,halogen, C₁₋₄ alkyl and C₁₋₄ haloalkyl, wherein the aliphatic portionsare optionally substituted with from one to three members selected fromthe group consisting of —OH, —OR^(o), —OC(O)NHR^(o), —OC(O)N(R^(o))₂,—SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o), —SO₂NH₂, —S(O)₂NHR^(o),—S(O)₂N(R^(o))₂, —NHS(O)₂R^(o), —NR^(o)S(O)₂R^(o), —C(O)NH₂,—C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o), —NHC(O)R^(o),—NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂, —NR^(o)C(O)NHR^(o),—NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂, —NHC(O)N(R^(o))₂, —CO₂H,—CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN, —NO₂, —NH₂, —NHR^(o),—N(R^(o))₂, —NR^(o)S(O)NH₂ and —NR^(o)S(O)₂NHR^(o), wherein each R^(o)is independently an unsubstituted C₁₋₆ alkyl. In certain instances ofthis embodiment, m is 0 or 1; R^(2a) is preferably hydrogen; andadditionally in other instances, R^(2c) is preferably selected from thegroup consisting of F, Cl, Br, CN, NO₂, —CO₂CH₃, —C(O)CH₃ and —S(O)₂CH₃.

In yet another embodiment, the R^(3a) substituent in formulae Ia and Ibis halogen, C₁₋₄ alkyl or C₁₋₄ haloalkyl.

In yet another embodiment, the R^(3a) moiety on the pyrazole ring informulae Ia and Ib is hydrogen, halogen, chloro, fluoro, bromo,oxazolyl, pyridyl, pyrimidinyl, imidazolyl, oxadiazolyl, thiazolyl, C₁₋₈alkyl, C₃₋₆ cycloalkyl or C₁₋₈ haloalkyl or cyano.

In yet another embodiment, the R^(3a) moiety on the pyrazole ring informulae Ia and Ib is hydrogen, halogen, chloro, fluoro, bromo,oxazolyl, pyridyl, pyrimidinyl, oxadiazolyl, thiazolyl, C₁₋₈ alkyl, C₃₋₆cycloalkyl or C₁₋₈ haloalkyl or cyano.

In a certain embodiment of the invention, in the compounds havingformulae Ia and Ib, R^(3a) is a member independently selected from thegroup consisting of hydrogen, halogen, —OR^(f), —NR^(f)R^(g),—C(O)R^(f), —C(O)OR^(f), —S(O)R^(f), —S(O)₂R^(f), —S(O)₃R^(f),—S(O)₃R^(h), —X³C(O)₂R^(f), X³S(O)₃R^(f), —S(O)₂NR^(f)R^(g),—X³S(O)₂NR^(f)R^(g), —R^(h), —CN, X³NR^(f)R^(g), NR^(g)C(O)R^(f), X³N₃and Y. The symbol Y is a five to six-membered aryl, a five orsix-membered heteroaryl ring or a three to eight-memberedheterocycloalkyl ring selected from the group consisting ofhomopiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl,piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl, piperazinzyl,phenyl, pyridyl, oxazolyl, pyrimidinyl, oxadiazolyl, imidazolyl,pyrazolyl, triazolyl and thiazolyl, optionally substituted with from oneto three substitutents selected from the group consisting of halogen,—OR^(f), —NR^(f)R^(g), —R^(h), —CN. Each R^(f) and R^(g) isindependently selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl andC₃₋₆ cycloalkyl, and each R^(h) is independently selected from the groupconsisting of C₁₋₆ alkyl, C₁₋₆ haloalkyl and C₃₋₆ cycloalkyl, whereinthe aliphatic portions of R^(f), R^(g) and R^(h) are optionally furthersubstituted with from one to three members selected from the groupconsisting of —OH, —OR^(o), —OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH,—SR^(o), —S(O)R^(o), —S(O)₂R^(o), —SO₂NH₂, —S(O)₂NHR^(o),—S(O)₂N(R^(o))₂, —NHS(O)₂R^(o), —NR^(o)S(O)₂R^(o), —C(O)NH₂,—C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o), —NHC(O)R^(o),—NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂, —NR^(o)C(O)NHR^(o),—NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂, —NHC(O)N(R^(o))₂, —CO₂H,—CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN, —NO₂, —NH₂, —NHR^(o),—N(R^(o))₂, —NR^(o)S(O)NH₂ and —NR^(o)S(O)₂NHR^(o), wherein R^(o) isunsubstituted C₁₋₆ alkyl.

In a certain embodiment of the invention, in the compounds havingformulae Ia and Ib, R^(3a) is a member independently selected from thegroup consisting of hydrogen, halogen, —OR^(f), —NR^(f)R^(g),—C(O)R^(f), —C(O)OR^(f), —S(O)R^(f), —S(O)₂R^(f), —S(O)₂NR^(f)R^(g),—R^(h), —CN, X³NR^(f)R^(g), NR^(g)C(O)R^(f), X³N₃ and —Y, wherein Y is afive to six-membered aryl, a five or six-membered heteroaryl ring or athree to eight-membered heterocycloalkyl ring selected from the groupconsisting of homopiperidinyl, morpholinyl, thiomorpholinyl,pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl,piperazinzyl, phenyl, pyridyl, oxazolyl, pyrimidinyl, oxadiazolyl andthiazolyl, optionally substituted with from one to three substitutentsselected from the group consisting of halogen, —OR^(f), —NR^(f)R^(g),—R^(h), —CN, wherein each R^(f) and R^(g) is independently selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl and C₃₋₆ cycloalkyl, and each R^(h)is independently selected from the group consisting of C₁₋₆ alkyl, C₁₋₆haloalkyl and C₃₋₆ cycloalkyl, wherein the aliphatic portions of R^(f),R^(g) and R^(h) are optionally further substituted with from one tothree members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein R^(o) is unsubstituted C₁₋₆ alkyl. Thesubscript m may be from 0 to 2; or alternatively from 0-1.

In another embodiment of the invention, in the compounds having formulaeIa or Ib, the symbol R^(3a) moiety on the pyrazole ring is hydrogen,halogen, chloro, fluoro, bromo, oxazolyl, pyridyl, oxadiazolylthiazolyl, —R^(h) or cyano; and optionally the symbol R¹, when present,is selected from the group consisting of —CO₂H or C₁₋₄ alkyl, optionallysubstituted with —OH, —OR^(m), —S(O)₂R^(m), —CO₂H and —CO₂R^(m). In yetanother embodiment, R¹, when present, is hydrogen or C₁₋₆ alkyl. m is aninteger from 0-2.

In another embodiment of the invention, in compounds of formulae Ia andIb, the R^(3a) substitutent is selected from the group consisting ofhydrogen, halogen, —OR^(f), NR^(f)R^(g), —R^(h), —Y, —CN, X³N₃,—SO₂R^(h), X³NR^(f)R^(g), X³Y, —S(O)₃R^(f), —C(C═NOR^(f))NR^(f)R^(g),—NO₂, and —NR^(g)C(O)R^(f), wherein Y is an optionally substituted groupselected from the group consisting of phenyl, pyridyl, pyrimidinyl,oxazolyl, thiazolyl, oxadiazolyl and morpholinyl, and R^(h) is anoptionally substituted group selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl and C₃₋₈ cycloalkyl, and R^(f) and R^(g) are eachindependently an optionally substituted group selected from the groupconsisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₈ cycloalkyl.In certain aspects of this embodiment, the R^(3a) substituent isselected from the group consisting of hydrogen, fluoro, chloro, bromo,iodo, amino, —CH₃, oxazolyl, thiazolyl, pyridyl, pyrimidinyl,morpholinyl, oxdiazolyl, —NHC(O)CH₃, —CN, CH₂N₃, CH₂SO₃H, NO₂,—(C═NOH)NH₂, —S(O)₂CH₃ and CH₂NH₂.

In yet another embodiment of the invention, in the compounds havingformulae Ia or Ib, the subscript m is 0 or 1; R^(2a) is hydrogen,halogen or —CN; R^(2c) is selected from the group consisting of F, Cl,Br, CN, NO₂, —CO₂CH₃, —C(O)CH₃ and —S(O)₂CH₃; R^(2d) is selected fromthe group consisting of —SR^(c), —O—X²—OR^(c), —X²—OR^(c), —R^(e) and—OR^(c); and R^(3a) substituents is selected from the group consistingof halogen, C₁₋₄ alkyl and C₁₋₄ haloalkyl, wherein the aliphaticportions of R^(3a) are optionally substituted with from one to threemembers selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein each R^(o) is independently anunsubstituted C₁₋₆ alkyl.

In one preferred embodiment, in the compounds of the invention havingthe formula Ib, when R^(2a) is H, R^(2c) is chloro, R^(2d) is methoxy, mis 0, a is N, c is N, and b and d are CH, then R^(3a) is other thanhydrogen, methyl, unsubstituted 2-pyridyl, unsubstituted 2-pyrimidinylor unsubstituted 2-oxazolyl.

In one specific embodiment, the present invention provides compoundshaving formula Ia and Ib wherein the subscript m is an integer of from 0to 4. The symbol R¹ is a substituent independently selected from thegroup consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl,—CO₂R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a) and —X¹OR^(a), —COR^(a),—CONR^(a)R^(b), —X¹NR^(a)R^(b), —X¹NR^(a)COR^(b), —X¹CONR^(a)R^(b),X¹S(O)₂NR^(a)R^(b) and X¹S(O)₂R^(a), wherein X¹ is C₁₋₄ alkylene andeach R^(a) and R^(b) is independently selected from the group consistingof hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl; and whereinthe aliphatic portions of each of said R¹ substituents is optionallysubstituted with from one to three members selected from the groupconsisting of —OH, —OR^(m), —OC(O)NHR^(m), —OC(O)N(R^(m))₂, —SH,—SR^(m), —S(O)R^(m), —S(O)₂R^(m), —SO₂NH₂, —S(O)₂NHR^(m),—S(O)₂N(R^(m))₂, —NHS(O)₂R^(m), —NR^(m)S(O)₂R^(m), —C(O)NH₂,—C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m), —NHC(O)R^(m),—NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂, —NR^(m)C(O)NHR^(m),—NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂, —NHC(O)N(R^(m))₂, —CO₂H,—CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN, —NO₂, —NH₂, —NHR^(m),—N(R^(m))₂, —NR^(m)S(O)NH₂ and —NR^(m)S(O)₂NHR^(m), wherein each R^(m)is independently an unsubstituted C₁₋₆ alkyl. The symbols R^(2a), R^(2c)and R^(2d) are each substituents independently selected from the groupconsisting of hydrogen, halogen, cyano, heteroaryl, —NO₂, —CO₂R^(c),—CONR^(c)R^(d), —C(O)R^(c), —S(O)R^(e), —S(O)₂R^(e), —C(NOR^(c))R^(d),—C(NR^(c)V)═NV, —N(V)C(R^(c))═NV, —X²C(NOR^(c))R^(d), —X²C(NR^(c)V)═NV,—X²N(V)C(R^(c))═NV, —X²NR^(c)R^(d), —X²SR^(c), —X²CN, —X²NO₂,—X²CO₂R^(c), —X²CONR^(c)R^(d), —X²C(O)R^(c), —X²OC(O)NR^(c)R^(d),—X²NR^(d)C(O)R^(c), —X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d),—X²NH—C(NH₂)═NH, —X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e),—X²NH—C(NHR^(e))═NH, —X²S(O)R^(e), —X²S(O)₂R^(e), —X²NR^(c)S(O)₂R^(e),—X²S(O)₂NR^(c)R^(d), —X²N₃, —OR^(c), —SR^(c), —NR^(d)C(O)R^(c),—NR^(d)C(O)₂R^(e), —S(O)₂R^(e), —S(O)₂NR^(c)R^(d), —X²OR^(c),—O—X²OR^(c), —X²NR^(c)R^(d), —O—X²NR^(c)R^(d) and —NR^(d)—X²CO₂R^(c).Within each of R^(2a), R^(2c) and R^(2d), X² is C₁₋₄ alkylene and eachR^(c) and R^(d) is independently selected from hydrogen, C₁₋₈ alkyl,C₁₋₈ haloalkyl, and C₃₋₆ cycloalkyl. Optionally, R^(c) and R^(d) whenattached to the same nitrogen atom can be combined with the nitrogenatom to form a five or six-membered ring having from 0 to 2 additionalheteroatoms as ring members. The symbol R^(e) is independently selectedfrom the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl and heteroaryl, and each ofR^(c), R^(d) and R^(e) is optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(n),—OC(O)NHR^(n), —OC(O)N(R^(n))₂, —SH, —SR^(n), —S(O)R^(n), —S(O)₂R^(n),—SO₂NH₂, —S(O)₂NHR^(n), —S(O)₂N(R^(n))₂, —NHS(O)₂R^(n),—NR^(n)S(O)₂R^(n), —C(O)NH₂, —C(O)NHR^(n), —C(O)N(R^(n))₂, —C(O)R^(n),—NHC(O)R^(n), —NR^(n)C(O)R^(n), —NHC(O)NH₂, —NR^(n)C(O)NH₂,—NR^(n)C(O)NHR^(n), —NHC(O)NHR^(n), —NR^(n)C(O)N(R^(n))₂,—NHC(O)N(R^(n))₂, —CO₂H, —CO₂R^(n), —NHCO₂R^(n), —NR^(n)CO₂R^(n), —CN,—NO₂, —NH₂, —NHR^(n), —N(R^(n))₂, —NR^(n)S(O)NH₂ and—NR^(n)S(O)₂NHR^(n), wherein each R^(n) is independently anunsubstituted C₁₋₆ alkyl; and wherein V is independently selected fromthe group consisting of —R^(c), —CN, —CO₂R^(e) and —NO₂. Each of ringvertices a, b, c and d in formulae Ia and Ib is independently selectedfrom N and C(R^(3a)), and from one to two of said ring vertices is N.The symbol R^(3a) in formulae Ia and Ib is independently selected fromthe group consisting of hydrogen, halogen, —OR^(f), —OC(O)R^(f),—NR^(f)R^(g), —SR^(f), —R^(h), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g),—C(O)R^(f), —OC(O)NR^(f)R^(g), —NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h),—NR^(f)—C(O)NR^(f)R^(g), —NH—C(NH₂)═NH, —NR^(h)C(NH₂)═NH,—NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH, —S(O)R^(h), —S(O)₂R^(h),—NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —NR^(f)S(O)₂R^(h),—NR^(f)S(O)₂NR^(f)R^(g), —N₃, —X³OR^(f), —X³OC(O)R^(f), —X³NR^(f)R^(g),—X³SR^(f), —X³CN, —X³NO₂, —X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f),—X³OC(O)NR^(f)R^(g), —X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h),—X³NR^(f)—C(O)NR^(f)R^(g), —X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH,—X³NH—C(NH₂)═NR^(h), —X³NH—C(NHR^(h))═NH, —X³S(O)R^(h), —X³S(O)₂R^(h),—X³NR^(f)S(O)₂R^(h), —X³S(O)₂NR^(f)R^(g), —Y, —X³Y and —X³N₃. The symbolY is a five to ten-membered aryl, heteroaryl or heterocycloalkyl ring,optionally substituted with from one to three substitutents selectedfrom the group consisting of halogen, —OR^(f), —NR^(f)R^(g), —R^(h),—SR^(f), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h),—S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g), —X³NR^(f)S(O)₂R^(h) and—X³S(O)₂NR^(f)R^(g), and wherein each X³ is independently selected fromthe group consisting of C₁₋₄ alkylene, C₂₋₄ alkenylene and C₂₋₄alkynylene; each R^(f) and R^(g) is independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄ alkyl, and aryloxy-C₁₋₄ alkyl,or when attached to the same nitrogen atom can be combined with thenitrogen atom to form a five or six-membered ring having from 0 to 2additional heteroatoms as ring members; and each R^(h) is independentlyselected from the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄alkyl, and aryloxy-C₁₋₄ alkyl, wherein the aliphatic portions of X³,R^(f), R^(g) and R^(h) are optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein R^(o) is unsubstituted C₁₋₆ alkyl.

In another specific embodiment, in compounds having formula Ia and Ib,R¹ is independently selected from the group consisting of C₁₋₈ alkyl,C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, —CO₂R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a),—X¹OR^(a), —COR^(a), —CONR^(a)R^(b), —X¹NR^(a)R^(b), —X¹NR^(a)COR^(b),—X¹CONR^(a)R^(b), X¹S(O)₂NR^(a)R^(b) and X¹S(O)₂R^(a), wherein X¹ isC₁₋₄ alkylene and each R^(a) and R^(b) is independently selected fromthe group consisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl. The aliphatic portions of each of said R¹ substituents isoptionally substituted with from one to three members selected from thegroup consisting of —OH, —OR^(m), —OC(O)NHR^(m), —OC(O)N(R^(m))₂, —SH,—SR^(m), —S(O)R^(m), —S(O)₂R^(m), —SO₂NH₂, —S(O)₂NHR^(m),—S(O)₂N(R^(m))₂, —NHS(O)₂R^(m), —NR^(m)S(O)₂R^(m), —C(O)NH₂,—C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m), —NHC(O)R^(m),—NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂, —NR^(m)C(O)NHR^(m),—NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂, —NHC(O)N(R^(m))₂, —CO₂H,—CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN, —NO₂, —NH₂, —NHR^(m),—N(R^(m))₂, —NR^(m)S(O)NH₂ and —NR^(m)S(O)₂NHR^(m), wherein each R^(m)is independently an unsubstituted C₁₋₆ alkyl. The substituents R^(2a),R^(2c) and R^(2d) are each independently selected from the groupconsisting of hydrogen, halogen, cyano, heteroaryl, —NO₂, —CO₂R^(c),—CONR^(c)R^(d), —C(O)R^(c), —S(O)R^(e), —S(O)₂R^(e), —R^(e),—C(NOR^(c))R^(d), —C(NR^(c)V)═NV, —N(V)C(R^(c))═NV, —X²C(NOR^(c))R^(d),—X²C(NR^(c)V)═NV, —X²N(V)C(R^(c))═NV, —X²NR^(c)R^(d), —X²SR^(c), —X²CN,—X²NO₂, —X²CO₂R^(c), —X²CONR^(c)R^(d), —X²C(O)R^(c),—X²OC(O)NR^(c)R^(d), —X²NR^(d)C(O)R^(c), —X²NR^(d)C(O)₂R^(e),—X²NR^(c)C(O)NR^(c)R^(d), —X²NH—C(NH₂)═NH, —X²NR^(e)C(NH₂)═NH,—X²NH—C(NH₂)═NR^(e), —X²NH—C(NHR^(e))═NH, —X²S(O)R^(e), —X²S(O)₂R^(e),—X²NR^(c)S(O)₂R^(e), —X²S(O)₂NR^(c)R^(d), —X²N₃, —OR^(c), —SR^(c),—R^(e), —NR^(d)C(O)R^(c), —NR^(d)C(O)₂R^(e), —S(O)₂R^(e),—S(O)₂NR^(c)R^(d), —X²OR^(c), —O—X²OR^(c), —X²NR^(c)R^(d),—O—X²NR^(c)R^(d) and —NR^(d)—X²CO₂R^(c); in which within each of R^(2a),R^(2c) and R^(2d), X² is C₁₋₄ alkylene and each R^(c) and R^(d) isindependently selected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, or optionally, R^(c) and R^(d) when attached to the samenitrogen atom can be combined with the nitrogen atom to form a five orsix-membered ring having from 0 to 2 additional heteroatoms as ringmembers; and each R^(e) is independently selected from the groupconsisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, aryl and heteroaryl, and each of R^(c), R^(d) and R^(e) isoptionally further substituted with from one to three members selectedfrom the group consisting of —OH, —OR^(n), —OC(O)NHR^(n),—OC(O)N(R^(n))₂, —SH, —SR^(n), —S(O)R^(n), —S(O)₂R^(n), —SO₂NH₂,—S(O)₂NHR^(n), —S(O)₂N(R^(n))₂, —NHS(O)₂R^(n), —NR^(n)S(O)₂R^(n),—C(O)NH₂, —C(O)NHR^(n), —C(O)N(R^(n))₂, —C(O)R^(n), —NHC(O)R^(n),—NR^(n)C(O)R^(n), —NHC(O)NH₂, —NR^(n)C(O)NH₂, —NR^(n)C(O)NHR^(n),—NHC(O)NHR^(n), —NR^(n)C(O)N(R^(n))₂, —NHC(O)N(R^(n))₂, —CO₂H,—CO₂R^(n), —NHCO₂R^(n), —NR^(n)CO₂R^(n), —CN, —NO₂, —NH₂, —NHR^(n),—N(R^(n))₂, —NR^(n)S(O)NH₂ and —NR^(n)S(O)₂NHR^(n). Each R^(n) isindependently an unsubstituted C₁₋₆ alkyl; and wherein V isindependently selected from the group consisting of —R^(c), —CN,—CO₂R^(e) and —NO₂. Each of ring vertices a, b, c and d in formulae Iaand Ib is independently selected from N and C(R^(3a)), and from one totwo of said ring vertices is N. The substituent R^(3a) is independentlyselected from the group consisting of hydrogen, halogen, —OR^(f),—OC(O)R^(f), —NR^(f)R^(g), —SR^(f), —R^(h), —CN, —NO₂, —CO₂R^(f),—CONR^(f)R^(g), —C(O)R^(f), —OC(O)NR^(f)R^(g), —NR^(g)C(O)R^(f),—NR^(g)C(O)₂R^(h), —NR^(f)—C(O)NR^(f)R^(g), —NH—C(NH₂)═NH,—NR^(h)C(NH₂)═NH, —NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH, —S(O)R^(h),—S(O)₂R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —NR^(f)S(O)₂R^(h),—NR^(f)S(O)₂NR^(f)R^(g), —N₃, —X³OR^(f), —X³OC(O)R^(f), —X³NR^(f)R^(g),—X³SR^(f), —X³CN, —X³NO₂, —X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f),—X³OC(O)NR^(f)R^(g), —X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h),—X³NR^(f)—C(O)NR^(f)R^(g), —X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH,—X³NH—C(NH₂)═NR^(h), —X³NH—C(NHR^(h))═NH, —X³S(O)R^(h), —X³S(O)₂R^(h),—X³NR^(f)S(O)₂R^(h), —X³S(O)₂NR^(f)R^(g), —Y, —X³Y and —X³N₃. The symbolY is a five to ten-membered aryl, heteroaryl or heterocycloalkyl ring,optionally substituted with from one to three substitutents selectedfrom the group consisting of halogen, —OR^(f), —NR^(f)R^(g), —R^(h),—SR^(f), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h),—S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g), —X³NR^(f)S(O)₂R^(h) and—X³S(O)₂NR^(f)R^(g), and wherein each X³ is independently selected fromthe group consisting of C₁₋₄ alkylene, C₂₋₄ alkenylene and C₂₋₄alkynylene and each R^(f) and R^(g) is independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄ alkyl, and aryloxy-C₁₋₄ alkyl,or when attached to the same nitrogen atom can be combined with thenitrogen atom to form a five or six-membered ring having from 0 to 2additional heteroatoms as ring members, and each R^(h) is independentlyselected from the group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, aryl, heteroaryl, aryl-C₁₋₄alkyl, and aryloxy-C₁₋₄ alkyl, wherein the aliphatic portions of X³,R^(f), R^(g) and R^(h) are optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R)₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R^(o))₂, —NR^(o)S(O)NH₂ and—NR^(o)S(O)₂NHR^(o), wherein R^(o) is unsubstituted C₁₋₆ alkyl.

In another specific embodiment of the invention, in compounds havingformula Ia or Ib, each R¹ is a substituent independently selected fromthe group consisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl,—CO₂R^(a), —X¹CO₂R^(a), —X¹SO₂R^(a) and —X¹OR^(a), wherein the aliphaticportions of each of said R¹ substituents is optionally substituted withfrom one to three members selected from the group consisting of —OH,—OR^(m), —OC(O)NHR^(m), —OC(O)N(R^(m))₂, —SH, —SR^(m), —S(O)R^(m),—S(O)₂R^(m), —SO₂NH₂, —S(O)₂NHR^(m), —S(O)₂N(R^(m))₂, —NHS(O)₂R^(m),—NR^(m)S(O)₂R^(m), —C(O)NH₂, —C(O)NHR^(m), —C(O)N(R^(m))₂, —C(O)R^(m),—NHC(O)R^(m), —NR^(m)C(O)R^(m), —NHC(O)NH₂, —NR^(m)C(O)NH₂,—NR^(m)C(O)NHR^(m), —NHC(O)NHR^(m), —NR^(m)C(O)N(R^(m))₂,—NHC(O)N(R^(m))₂, —CO₂H, —CO₂R^(m), —NHCO₂R^(m), —NR^(m)CO₂R^(m), —CN,—NO₂, —NH₂, —NHR^(m), —N(R^(m))₂, —NR^(m)S(O)NH₂ and—NR^(m)S(O)₂NHR^(m), wherein each R^(m) is independently anunsubstituted C₁₋₆ alkyl. The R^(2a) substituent is selected from thegroup consisting of hydrogen, halogen, cyano, heteroaryl, —NO₂,—CO₂R^(c), —CONR^(c)R^(d), —C(O)R^(o), —S(O)R^(e), —S(O)₂R^(e), —R^(e),—C(NOR^(c))R^(d), —C(NR^(c)V)═NV, —N(V)C(R^(c))═NV, —X²C(NOR^(c))R^(d),—X²C(NR^(c)V)═NV, —X²N(V)C(R^(c))═NV, —X²NR^(c)R^(d), —X²SR^(c), —X²CN,—X²NO₂, —X²CO₂R^(c), —X²CONR^(c)R^(d), —X²C(O)R^(c),—X²OC(O)NR^(c)R^(d), —X²NR^(d)C(O)R^(c), —X²NR^(d)C(O)₂R^(e),—X²NR^(c)C(O)NR^(c)R^(d), —X²NH—C(NH₂)═NH, —X²NR^(e)C(NH₂)═NH,—X²NH—C(NH₂)═NR^(e), —X²NH—C(NHR^(e))═NH, —X²S(O)R^(e), —X²S(O)₂R^(e),—X²NR^(c)S(O)₂R^(e), —X²S(O)₂NR^(c)R^(d) and —X²N₃. The R^(2c) andR^(2d) substituents are each independently selected from the groupconsisting of halogen, —OR^(c), —SR^(c), —R^(e), —CN, —NO₂, —CO₂R^(c),—C(O)R^(c), —NR^(d)C(O)R^(c), —NR^(d)C(O)₂R^(e), —S(O)₂R^(e),—S(O)₂NR^(c)R^(d), —X²OR^(c), —O—X²OR^(c), —X²NR^(c)R^(d),—O—X²NR^(c)R^(d) and —NR^(d)—X²CO₂R^(c). Each R^(3a) substituent isindependently selected from the group consisting of hydrogen, halogen,—OR^(f), —OC(O)R^(f), —NR^(f)R^(g), —SR^(f), —R^(h), —CN, —NO₂,—CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f), —OC(O)NR^(f)R^(g),—NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h), —NR^(f)—C(O)NR^(f)R^(g),—S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g),—NR^(f)S(O)₂NR^(f)R^(g), —X³OR^(f), —X³NR^(f)R^(g), —X³SR^(f), —X³CN,—C(C═NOR^(f))NR^(f)R^(g), X³SO₃R^(f), —X³CO₂R^(f), —X³CONR^(f)R^(g),—X³C(O)R^(f), —X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h), —Y, —X³Y, —X³N₃,wherein Y is selected from the group consisting of a five orsix-membered aryl ring, a five or six-membered heteroaryl ring and threeto eight membered heterocycloalkyl ring, wherein said Y group isoptionally substituted with from one to three substitutents selectedfrom the group consisting of halogen, —OR^(f), —NR^(f)R^(g), —R^(h),—SR^(f), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—NR^(g)C(O)R^(f), —S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h) and—S(O)₂NR^(f)R^(g), and wherein each X³ is independently C₁₋₄ alkylene,and each R^(f) and R^(g) is independently selected from hydrogen, C₁₋₈alkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl, and each R^(h) isindependently selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl and C₃₋₆ cycloalkyl, wherein the aliphatic portions of X³,R^(f), R^(g) and R^(h) is optionally further substituted with from oneto three members selected from the group consisting of —OH, —OR^(o),—OC(O)NHR^(o), —OC(O)N(R^(o))₂, —SH, —SR^(o), —S(O)R^(o), —S(O)₂R^(o),—SO₂NH₂, —S(O)₂NHR^(o), —S(O)₂N(R^(o))₂, —NHS(O)₂R^(o),—NR^(o)S(O)₂R^(o), —C(O)NH₂, —C(O)NHR^(o), —C(O)N(R^(o))₂, —C(O)R^(o),—NHC(O)R^(o), —NR^(o)C(O)R^(o), —NHC(O)NH₂, —NR^(o)C(O)NH₂,—NR^(o)C(O)NHR^(o), —NHC(O)NHR^(o), —NR^(o)C(O)N(R^(o))₂,—NHC(O)N(R^(o))₂, —CO₂H, —CO₂R^(o), —NHCO₂R^(o), —NR^(o)CO₂R^(o), —CN,—NO₂, —NH₂, —NHR^(o), —N(R)₂, —NR^(o)S(O)NH₂ and —NR^(o)S(O)₂NHR^(o),wherein each R^(o) is independently an unsubstituted C₁₋₆ alkyl.

In another embodiment of the invention, the compounds of the inventionhaving formula Ib is represented by formulae Ib¹ and Ib²:

or a pharmaceutically acceptable salt, hydrate or N-oxide thereof. Inone embodiment, in formulae Ib¹ and Ib², R^(2c) is halogen, cyano ornitro; the symbol R^(2d) is selected from —SR^(c), —O—X²—OR^(c),—X²—OR^(c), R^(e), —OR^(c) and —NR^(d)C(O)R^(c); each of ring verticesa, b, c and d is independently selected from N and C(R^(3a)), and fromone to two of said ring vertices is N; and each R^(3a) is independentlyselected from the group consisting of hydrogen, halogen, C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆ heterocycloalkyl, —S(O)₂R^(h),amino, phenyl, pyridyl, pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolylimidazolyl and thiazolyl. In one embodiment, the ring vertex a is N. Inanother embodiment, the ring vertex b is N. In another embodiment, thering vertex c is N. In another embodiment, the ring vertex d is N. Inyet another embodiment, the ring vertices a and c are each N; b ishydrogen; and d is C(R^(3a)), wherein R^(3a) on ring vertex d is otherthan hydrogen. In another embodiment, the ring vertex a is N; b isC(R^(3a)) wherein R^(3a) on ring vertex b is other than hydrogen; and cand d are each hydrogen. In another embodiment, the ring vertex a is N;b and c are each hydrogen; and d is C(R^(3a)), wherein R^(3a) on ringvertex d is other than hydrogen. In another embodiment, the ring vertexa is C(R^(3a)), wherein R^(3a) on ring vertex a is other than hydrogen;b is N; c and d are each hydrogen. In another embodiment, the ringvertex a is N; b and d are each hydrogen; and c is C(R^(3a)); whereinR^(3a) on ring vertex c is other than hydrogen. In another embodiment,the ring vertices a and c are each N; b is hydrogen; and d is C(R^(3a)),wherein R^(3a) on ring vertex d is other than hydrogen.

In yet another embodiment of the invention, in formulae Ib¹ and Ib²,R^(2c) and R^(2d) are each a member independently selected from thegroup consisting of halogen, C₁₋₄ alkoxy, C₁₋₄ alkyl and C₁₋₄ haloalkyl;R^(3a) on the pyrazole ring is an imidazolyl group optionallysubstituted with C₁₋₄ alkyl; and the ring vertex a is nitrogen and thering vertices b, c, and d are each C(H). Within certain aspects of thisembodiment, R^(2c) is selected from the group consisting of fluoro,chloro, bromo and iodo; and R^(2d) is selected from the group consistingof methoxy, ethoxy, trifluoromethoxy, methyl, ethyl, trifluoromethyl and2-fluoroethoxy.

In yet another embodiment of the invention, the compounds of theinvention having formula Ib is represented by formulae Ib³ and Ib⁴:

or a pharmaceutically acceptable salt, hydrate and an N-oxide thereof.In one embodiment, in formulae Ib³ and Ib⁴, R^(2c) is independentlyhalogen, cyano or nitro; R^(2d) is selected from —SR^(c), —O—X²—OR^(c),—X²—OR^(c), R^(e), —OR^(c), —NR^(c)R^(d), —NR^(c)S(O)₂R^(e) and—NR^(d)C(O)R^(c); R^(2a) is selected from the group consisting of F, Cl,Br, I, —CO₂Me, —CONH₂, CN, oxazolyl, —CH₂NH₂, —CH₂NHMe, —CH₂NMe₂ and−CH═N—OH; each of ring vertices a, b, c and d is independently selectedfrom N and C(R^(3a)), and from one to two of said ring vertices is N;and each R^(3a) is independently selected from the group consisting ofhydrogen, halogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆heterocycloalkyl, —S(O)₂R^(h), amino, phenyl, pyridyl, pyrimidinyl,oxazolyl, oxadiazolyl, isoxazolyl and thiazolyl. In one embodiment thering vertex a is N. In another embodiment the ring vertex b is N. Inanother embodiment the ring vertex c is N. In another embodiment thering vertex d is N. In yet another embodiment, the ring vertices a and care each N; b is hydrogen; and d is C(R^(3a)), wherein R^(3a) on ringvertex d is other than hydrogen. In another embodiment, the ring vertexa is N; b is C(R^(3a)) wherein R^(3a) on ring vertex b is other thanhydrogen; and c and d are each hydrogen. In another embodiment, the ringvertex a is N; b and c are each hydrogen; and d is C(R^(3a)), whereinR^(3a) on ring vertex d is other than hydrogen. In another embodiment,the ring vertex a is C(R^(3a)), wherein R^(3a) on ring vertex a is otherthan hydrogen; b is N; c and d are each hydrogen. In another embodiment,the ring vertex a is N; b and d are each hydrogen; and c is C(R^(3a));wherein R^(3a) on ring vertex c is other than hydrogen. In anotherembodiment, the ring vertices a and c are each N; b is hydrogen; and dis C(R^(3a)), wherein R^(3a) on ring vertex d is other than hydrogen.

In yet another embodiment of the invention, in formulae Ib³ and Ib⁴,R^(2a), R^(2c) and R^(2d) are each a member independently selected fromthe group consisting of hydrogen, halogen, C₁₋₄ alkoxy, C₁₋₄ alkyl andC₁₋₄ haloalkyl; R^(3a) on the pyrazole ring is an imidazolyl groupoptionally substituted with C₁₋₄ alkyl; and the ring vertex a isnitrogen and the ring vertices b, c, and d are each C(H). Within certainaspects of this embodiment, R^(2a) and R^(2c) in are each independentlyselected from the group consisting of fluoro, chloro, bromo and iodo;and R^(2d) is selected from the group consisting of methoxy, ethoxy,trifluoromethoxy, methyl, ethyl, trifluoromethyl and 2-fluoroethoxy.

In yet another embodiment, the compounds of the invention having formulaIa is represented by formulae Ia¹ or Ia²:

or a pharmaceutically acceptable salt, hydrate or N-oxide thereof;wherein the symbol R^(2c) is halogen, cyano or nitro; the symbol R^(2d)is selected from —SR^(c), —O—X²—OR^(c), —X²—OR^(c), —R^(e), —OR^(c) and—NR^(d)C(O)R^(c); each of ring vertices a, b, c and d is independentlyselected from N and C(R^(3a)), and from one to two of said ring verticesis N; and each R^(3a) is independently selected from the groupconsisting of hydrogen, halogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₆cycloalkyl, C₃₋₆ heterocycloalkyl, —S(O)₂R^(h), amino, phenyl, pyridyl,pyrimidinyl, oxazoylyl, oxadiazolyl, isoxazolyl and thiazolyl. In oneembodiment, the ring vertex d is N. In another embodiment the ringvertex b is N. In another embodiment the ring vertex c is N. In anotherembodiment the ring vertex d is N. In another embodiment, the ringvertex a is N; b and d are each hydrogen; and c is C(R^(3a)) whereinR^(3a) is other than hydrogen. In another embodiment, the ring vertex ais C(R^(3a)), wherein R^(3a) on ring vertex a is other than hydrogen; bis N; and c and d are each hydrogen. In another embodiment, the ringvertex a is N; b and c are each hydrogen; and d is C(R^(3a)) whereinR^(3a) on ring vertex d is other than hydrogen. In another embodiment,the ring vertex a is C(R^(3a)), wherein R^(3a) on ring vertex a is otherthan hydrogen; b and c are each hydrogen; and d is N. In anotherembodiment, the ring vertex a is C(R^(3a)), wherein R^(3a) on ringvertex a is other than hydrogen; b and d are each N; and c is hydrogen.In another embodiment, the ring vertices a and b are each hydrogen; c isC(R^(3a)), wherein R^(3a) on ring vertex c is other than hydrogen; and dis N.

In yet another embodiment, the compounds of the invention having formulaIa is represented by formula Ia³ and Ia⁴:

or pharmaceutically acceptable salt, hydrate or N-oxide thereof; whereinR^(2c) is halogen, cyano or nitro; R^(2d) is selected from —SR^(c),—O—X²—OR^(c), —X²—OR^(c), —R^(e), —OR^(c) and —NR^(d)C(O)R^(c); R^(2a)is selected from the group consisting of F, Cl, Br, I, —CO₂Me, —CONH₂,CN, oxazolyl, —CH₂NH₂, —CH₂NHMe, and —CH₂NMe₂; each of ring vertices a,b, c and d is independently selected from N and C(R^(3a)), and from oneto two of said ring vertices is N; and each R^(3a) is independentlyselected from the group consisting of hydrogen, halogen, C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆ heterocycloalkyl, —S(O)₂R^(h),amino, phenyl, pyridyl, pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolyland thiazolyl. In one embodiment, the ring vertex a is N. In anotherembodiment the ring vertex b is N. In another embodiment the ring vertexc is N. In another embodiment, the ring vertex d is N. In anotherembodiment, the ring vertex a is C(R^(3a)), wherein R^(3a) on ringvertex a is other than hydrogen; b is N; and c and d are each hydrogen.In another embodiment, the ring vertex a is N; b and c are eachhydrogen; and d is C(R^(3a)) wherein R^(3a) on ring vertex d is otherthan hydrogen. In another embodiment, the ring vertex a is C(R^(3a)),wherein R^(3a) on ring vertex a is other than hydrogen; b and c are eachhydrogen; and d is N. In another embodiment, the ring vertex a isC(R^(3a)), wherein R^(3a) on ring vertex a is other than hydrogen; b andd are each N; and c is hydrogen. In another embodiment, the ringvertices a and b are each hydrogen; c is C(R^(3a)), wherein R^(3a) onring vertex c is other than hydrogen; and d is N.

A family of specific compound of particular interest having formulae Iaand Ib consists of compounds, pharmaceutically acceptable salts,hydrates or N-oxides thereof, as set forth in Table 1.

TABLE 1 1.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyridin-1-yl-ethanone 2.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyridin-2-yl-ethanone 3.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4-b]pyridin-2-yl)-ethanone 4.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyrazin-1-yl-7-oxide)-ethanone 5.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyrazin-1-yl-7-oxide)-ethanone 6.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone. 7.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone. 8.2-(3-Amino-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-ethanone. 9.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 10.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 11.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone. 12.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone. 13.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[4,3-c]pyridin-1-yl-ethanone. 14.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-2-yl-ethanone. 15.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 16.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 17.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone. 18.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone. 19.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 20.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 21.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-fluoro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 22.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 23.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-2-yl-ethanone 24.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-1-yl-ethanone 25.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone 26.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 27.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 28.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 29.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 30.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 31.1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile 32.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyridin-1-yl-2-oxide)-ethanone 33.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 34.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-b]pyridin-2-yl)-ethanone 35.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-morpholin-4-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 36.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-1-yl-ethanone 37.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-c]pyridin-1-yl-6-oxide)-ethanone 38.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[4,3-c]pyridin-2-yl)-ethanone 39.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[3,4-b]pyridin-2-yl)-ethanone 40.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 41.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methanesulfonyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 42.2-(3-Azidomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 43.(1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridin-3-yl)-methanesulfonic acid 44.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 45.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidin-2-yl)-ethanone 46.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidin-1-yl)-ethanone 47.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methoxy-pyrazolo[3,4-d]pyrimidin-1-yl)-ethanone 48.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridin-2-yl)-ethanone 49.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 50.2-(6-Azido-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 51.2-(6-Amino-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 52.2-(7-Azido-pyrazolo[3,4-c]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 53.2-(7-Amino-pyrazolo[3,4-c]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 54.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-2-yl)-ethanone 55.2-(5-Amino-3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 56.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-5-nitro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 57.2-(3-Amino-6-methyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 58.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 59.1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine 60.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 61.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(5-methy1-[1,2,4]oxadiazol-3-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 62.N-(1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridin-6-yl)-acetamide 63.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methanesulfonyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 64.2-(3-Aminomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 65.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 66.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 67.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 68.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 69.1-[4-(4-Chloro-3-ethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 70.1-{4-[4-Chloro-3-(2-fluoro-ethoxy)-phenyl]-piperazin-1-yl}-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 71.1-[4-(4-Chloro-3-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 72.1-[4-(4-Chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 73.1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 74.1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 75.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 76.1-{4-[4-Chloro-3-(2-fluoro-ethoxy)-phenyl]-2-methyl-piperazin-1-yl}-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 77.1-[4-(4-Chloro-3-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 78.1-[4-(4-Chloro-5-ethoxy-2-fluoro-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 79.1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 80.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 81.1-[4-(4-Chloro-3-ethoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 82.1-{4-[4-Chloro-2-fluoro-5-(2-fluoro-ethoxy)-phenyl]-piperazin-1-yl}-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 83.1-[4-(4-Chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 84.1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 85.1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 86.2-[4-Amino-3-(1H-imidazol-2-yl)-pyrazolo[3,4-d]pyrimidin-1-yl]-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 87.2-[5-Chloro-3-(1H-imidazol-2-yl)-indazol-1-yl]-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone 88.2-[6-Chloro-3-(1H-imidazol-2-yl)-indazol-1-yl]-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone

Another family of specific compounds of particular interest havingformulae Ia and Ib consists of compounds, pharmaceutically acceptablesalts, hydrates or N-oxides thereof as set forth in Table 2.

Preparation of Compounds

As provided in the examples below, the compounds and intermediates ofthe present invention can be prepared by one of skill in the art in acomponent assembly manner. Schemes 1A-1M illustrate a variety of methodsfor the preparation of a variety of azaindazole-type derivatives. Ineach of these schemes, X is halogen; Nu is nucleophilic group; thesymbol {circle around (N)} within an aryl ring indicate the replacementof one to two carbon(s) of said aryl ring vertex (vertices) withnitrogen atom(s); L is a ligand; and non-interferring substituents areprovided as —R, —R′, —R″, and —R′″.

Scheme 1A

Scheme 1A shows the synthesis of azaindazole derivatives fromhalo-pyridine-carbaldehyde or ketone.

Scheme 1B

Scheme 1B shows the synthesis of azaindazole derivatives fromhalo-cyanopyridines.

Scheme 1C

Scheme 1C shows the synthesis of azaindazole derivatives fromamino-methyl-pyridine.

Scheme 1D

Scheme 1D shows the reaction of azaindazole derivatives with anα-haloacetate or α-haloacetamide.

Scheme 1E

Scheme 1E shows the reaction of azaindazole derivatives with anelectrophilic halogen source (X+).

Scheme 1F

Scheme 1F shows a metal-assisted coupling reaction of a halo-azaindazolederivative.

Scheme 1G

Scheme 1G shows a metal-assisted amination reaction of ahalo-azaindazole derivative.

Scheme 1H

Scheme 1H shows the amination of an azaindazole derivative.

Scheme 1I

Scheme 1I shows the functionalization of an azaindazole derivative.

Scheme 1J

Scheme IJ shows the synthesis of a pyrazolopyrazine derivative.

Scheme 1K

Scheme IK shows the synthesis of a pyrazolopyrimidine derivative.

Scheme 1L

Scheme IL shows the synthesis of a pyrazolopyrimidine derivative.

Scheme 1M

Scheme 1M shows the synthesis of N-oxide derivatives of the invention.

Scheme 1N

Scheme 1N shows the synthesis of 3-imidazolyl-azaindazoles derivatives.

Scheme 1N illustrates the synthesis of 3-imidazolyl substitutedpyrazolo[3,4-b]pyridines wherein R represents a non-interferringsubstituent, such as, for example, a protecting group, a carboxy esteror the remainder of the compound of formula Ib. As shown in Scheme 1N,the reaction of NH₂OH with 3-cyano pyrazolo[3,4-b]pyridine provides ahydroxylamidine compound, which upon reduction using hydrogen gas and acatalyst (e.g., Pd/C or Pd(OH)₂ produces an the amidine product.Cyclization of the amindine by treatment with chloroacetaldehyde willproduce the imidazole product.

Scheme 1O

Scheme 1O shows the synthesis of 3-imidazolyl-azaindazoles derivatives.

Scheme 1O illustrates the synthesis of 3-imidazolyl substitutedpyrazolo[3,4-b]pyridines. R′ represents a non-interfering substituent,such as, for example, a protecting group, a carboxy ester, or theremainder of the compound of formula Ib. As shown in Scheme 1O, thereaction of 3-cyano-pyrazolo[3,4-b]pyridine with ethylene diamineproduces the cyclized imidazoline product, which upon oxidation producesthe imidazole.

Scheme 1P

Scheme 1P shows the synthesis of 3-imidazolyl-azaindazoles derivatives.3-imidazolyl-azaindazoles shown in Scheme 1P can be produced in kilogramscale using the outlined synthetic route.

Scheme 1P illustrates the synthesis of 3-imidazolyl substitutedpyrazolo[3,4-b]pyridines, wherein R′ represents a non-interferingsubstituent, such as, for example, a protecting group, or a carboxyester, or the remainder of the compound of formula I (see also theExamples). As shown in Scheme 1P, using a transmetallation process, a3-iodo-pyrazolo[3,4-b]pyridine intermediate can be converted to a3-formyl-pyrazolo[3,4-b]pyridine intermediate, which upon treatment withglyoxal, is cyclized to form 3-imidazolyl-pyrazolo[3,4]pyridine product.

IV. Pharmaceutical Compositions

In addition to the compounds provided above, compositions for modulatingCCR1 activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self emulsifications as described in U.S. PatentApplication 2002-0012680, hard or soft capsules, syrups, elixirs,solutions, buccal patch, oral gel, chewing gum, chewable tablets,effervescent powder and effervescent tablets. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents, antioxidants andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as cellulose, silicon dioxide, aluminumoxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example PVP, cellulose, PEG, starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated,enterically or otherwise, by known techniques to delay disintegrationand absorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate may beemployed. They may also be coated by the techniques described in theU.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols. Additionally, the compounds can be administeredvia ocular delivery by means of solutions or ointments. Still further,transdermal delivery of the subject compounds can be accomplished bymeans of iontophoretic patches and the like. For topical use, creams,ointments, jellies, solutions or suspensions, etc., containing thecompounds of the present invention are employed. As used herein, topicalapplication is also meant to include the use of mouth washes andgargles.

In exemplary embodiment, the inhibitory agent of this invention may bedeposited within a medical device, such as a stent, and delivered to thetreatment site for treatment of a portion of the body.

Stents have been used as delivery vehicles for therapeutic agents (i.e.,drugs). Intravascular stents are generally permanently implanted incoronary or peripheral vessels. Stent designs include those of U.S. Pat.Nos. 4,733,655 (Palmaz), 4,800,882 (Gianturco), or 4,886,062 (Wiktor).Such designs include both metal and polymeric stents, as well asself-expanding and balloon-expandable stents. Stents may also used todeliver a drug at the site of contact with the vasculature, as disclosedin U.S. Pat. No. 5,102,417 (Palmaz) and in International PatentApplication Nos. WO 91/12779 (Medtronic, Inc.) and WO 90/13332(Cedars-Sanai Medical Center), U.S. Pat. No. 5,419,760 (Narciso, Jr.)and U.S. Pat. No. 5,429,634 (Narciso, Jr.), for example. Stents havealso been used to deliver viruses to the wall of a lumen for genedelivery, as disclosed in U.S. patent application Ser. No. 08/746,404,filed Nov. 8, 1996 (Donovan et al.).

The term “deposited” means that the inhibitory agent is coated,adsorbed, placed, or otherwise incorporated into the device by methodsknown in the art. For example, the inhibitory agent may be embedded andreleased from within (“matrix type”) or surrounded by and releasedthrough (“reservoir type”) polymer materials that coat or span themedical device. In the later example, the inhibitory agent may beentrapped within the polymer materials or coupled to the polymermaterials using one or more the techniques for generating such materialsknown in the art. In other formulations, the inhibitory agent may belinked to the surface of the medical device without the need for acoating by means of detachable bonds and release with time, can beremoved by active mechanical or chemical processes, or are in apermanently immobilized form that presents the inhibitory agent at theimplantation site.

In one embodiment, the inhibitory agent may be incorporated with polymercompositions during the formation of biocompatible coatings for medicaldevices, such as stents. The coatings produced from these components aretypically homogeneous and are useful for coating a number of devicesdesigned for implantation.

The polymer may be either a biostable or a bioabsorbable polymerdepending on the desired rate of release or the desired degree ofpolymer stability, but a bioabsorbable polymer is preferred for thisembodiment since, unlike a biostable polymer, it will not be presentlong after implantation to cause any adverse, chronic local response.Bioabsorbable polymers that could be used include, but are not limitedto, poly(L-lactic acid), polycaprolactone, polyglycolide (PGA),poly(lactide-co-glycolide) (PLLA/PGA), poly(hydroxybutyrate),poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester,polyanhydride, poly(glycolic acid), poly(D-lactic acid), poly(L-lacticacid), poly(D,L-lactic acid), poly(D,L-lactide) (PLA), poly(L-lactide)(PLLA), poly(glycolic acid-co-trimethylene carbonate) (PGA/PTMC),polyethylene oxide (PEO), polydioxanone (PDS), polyphosphoester,polyphosphoester urethane, poly(amino acids), cyanoacrylates,poly(trimethylene carbonate), poly(iminocarbonate), copoly(ether-esters)(e.g., PEO/PLA), polyalkylene oxalates, polyphosphazenes andbiomolecules such as fibrin, fibrinogen, cellulose, starch, collagen andhyaluronic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates,cross linked or amphipathic block copolymers of hydrogels, and othersuitable bioabsorbable polymers known in the art. Also, biostablepolymers with a relatively low chronic tissue response such aspolyurethanes, silicones, and polyesters could be used and otherpolymers could also be used if they can be dissolved and cured orpolymerized on the medical device such as polyolefins, polyisobutyleneand ethylene-alphaolefin copolymers; acrylic polymers and copolymers,vinyl halide polymers and copolymers, such as polyvinyl chloride;polyvinylpyrrolidone; polyvinyl ethers, such as polyvinyl methyl ether;polyvinylidene halides, such as polyvinylidene fluoride andpolyvinylidene chloride; polyacrylonitrile, polyvinyl ketones; polyvinylaromatics, such as polystyrene, polyvinyl esters, such as polyvinylacetate; copolymers of vinyl monomers with each other and olefins, suchas ethylene-methyl methacrylate copolymers, acrylonitrile-styrenecopolymers, ABS resins, and ethylene-vinyl acetate copolymers; pyrancopolymer; polyhydroxy-propyl-methacrylamide-phenol;polyhydroxyethyl-aspartamide-phenol; polyethyleneoxide-polylysinesubstituted with palmitoyl residues; polyamides, such as Nylon 66 andpolycaprolactam; alkyd resins, polycarbonates; polyoxymethylenes;polyimides; polyethers; epoxy resins, polyurethanes; rayon;rayon-triacetate; cellulose, cellulose acetate, cellulose butyrate;cellulose acetate butyrate; cellophane; cellulose nitrate; cellulosepropionate; cellulose ethers; and carboxymethyl cellulose.

Polymers and semipermeable polymer matrices may be formed into shapedarticles, such as valves, stents, tubing, prostheses and the like.

In one embodiment of the invention, the inhibitory agent of theinvention is coupled to a polymer or semipermeable polymer matrix thatis formed as a stent or stent-graft device.

Typically, polymers are applied to the surface of an implantable deviceby spin coating, dipping or spraying. Additional methods known in theart can also be utilized for this purpose. Methods of spraying includetraditional methods as well as microdeposition techniques with an inkjettype of dispenser. Additionally, a polymer can be deposited on animplantable device using photo-patterning to place the polymer on onlyspecific portions of the device. This coating of the device provides auniform layer around the device which allows for improved diffusion ofvarious analytes through the device coating.

In preferred embodiments of the invention, the inhibitory agent isformulated for release from the polymer coating into the environment inwhich the medical device is placed. Preferably, the inhibitory agent isreleased in a controlled manner over an extended time frame (e.g.,months) using at least one of several well-known techniques involvingpolymer carriers or layers to control elution. Some of these techniqueswere previously described in U.S. Patent Application 20040243225A1, theentire disclosure of which is incorporated in its entirety.

Moreover, as described for example in U.S. Pat. No. 6,770,729, which isincorporated herein in its entirety, the reagents and reactionconditions of the polymer compositions can be manipulated so that therelease of the inhibitory agent from the polymer coating can becontrolled. For example, the diffusion coefficient of the one or morepolymer coatings can be modulated to control the release of theinhibitory agent from the polymer coating. In a variation on this theme,the diffusion coefficient of the one or more polymer coatings can becontrolled to modulate the ability of an analyte that is present in theenvironment in which the medical device is placed (e.g. an analyte thatfacilitates the breakdown or hydrolysis of some portion of the polymer)to access one or more components within the polymer composition (and forexample, thereby modulate the release of the inhibitory agent from thepolymer coating). Yet another embodiment of the invention includes adevice having a plurality of polymer coatings, each having a pluralityof diffusion coefficients. In such embodiments of the invention, therelease of the inhibitory agent from the polymer coating can bemodulated by the plurality of polymer coatings.

In yet another embodiment of the invention, the release of theinhibitory agent from the polymer coating is controlled by modulatingone or more of the properties of the polymer composition, such as thepresence of one or more endogenous or exogenous compounds, oralternatively, the pH of the polymer composition. For example, certainpolymer compositions can be designed to release a inhibitory agent inresponse to a decrease in the pH of the polymer composition.Alternatively, certain polymer compositions can be designed to releasethe inhibitory agent in response to the presence of hydrogen peroxide.

V. Methods of Treating Diseases Modulated by CCR1, CCR2 and/or CCR3

In yet another aspect, the present invention provides methods oftreating CCR1, CCR2 and/or CCR3-mediated conditions or diseases byadministering to a subject having such a disease or condition, atherapeutically effective amount of a compound of formula I above. The“subject” is defined herein to include animals such as mammals,including, but not limited to, primates (e.g., humans), cows, sheep,goats, horses, dogs, cats, rabbits, rats, mice and the like.

CCR1 provides a target for interfering with or promoting specificaspects of immune cell functions, or more generally, with functionsassociated with CCR1 expression on a wide range of cell types in amammal, such as a human. Compounds that inhibit CCR1, are particularlyuseful for modulating monocyte, macrophage, lymphocyte, granulocyte, NKcell, mast cells, dendritic cell, neutrophils, and certain immunederived cell (for example, osteoclasts) function for therapeuticpurposes. Accordingly, the present invention is directed to compoundswhich are useful in the prevention and/or treatment of a wide variety ofinflammatory and immunoregulatory disorders and diseases (see Saeki, etal., Current Pharmaceutical Design 9:1201-1208 (2003)).

For example, an instant compound that inhibits one or more functions ofCCR1 may be administered to inhibit (i.e., reduce or prevent)inflammation or cellular infiltration associated with an immunedisorder. As a result, one or more inflammatory processes, such asleukocyte emigration or infiltration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, can be inhibited.For example, monocyte infiltration to an inflammatory site (e.g., anaffected joint in arthritis, or into the CNS in MS) can be inhibitedaccording to the present method.

Similarly, an instant compound that promotes one or more functions ofCCR1 is administered to stimulate (induce or enhance) an inflammatoryresponse, such as leukocyte emigration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, resulting in thebeneficial stimulation of inflammatory processes. For example, monocytescan be recruited to combat bacterial infections.

Diseases and conditions associated with inflammation, immune disordersand infection can be treated using the method of the present invention.In a preferred embodiment, the disease or condition is one in which theactions of immune cells such monocyte, macrophage, lymphocyte,granulocyte, NK cell, mast cell, dendritic cell, or certain immunederived cell (for example, osteoclasts) are to be inhibited or promoted,in order to modulate the inflammatory or autoimmune response.

In one group of embodiments, diseases or conditions, including chronicdiseases, of humans or other species can treated with modulators ofCCR1, CCR2 or CCR3 function. These diseases or conditions include: (1)allergic diseases such as systemic anaphylaxis or hypersensitivityresponses, drug allergies, insect sting allergies and food allergies,(2) inflammatory bowel diseases, such as Crohn's disease, ulcerativecolitis, ileitis and enteritis, (3) vaginitis, (4) psoriasis andinflammatory dermatoses such as dermatitis, eczema, atopic dermatitis,allergic contact dermatitis, urticaria and pruritus, (5) vasculitis, (6)spondyloarthropathies, (7) scleroderma, (8) asthma and respiratoryallergic diseases such as allergic asthma, allergic rhinitis,hypersensitivity lung diseases and the like, (9) autoimmune diseases,such as fibromyalagia, scleroderma, ankylosing spondylitis, juvenile RA,Still's disease, polyarticular juvenile RA, pauciarticular juvenile RA,polymyalgia rheumatica, rheumatoid arthritis, psoriatic arthritis,osteoarthritis, polyarticular arthritis, multiple sclerosis, systemiclupus erythematosus, type I diabetes, type II diabetes,glomerulonephritis, and the like, (10) graft rejection (includingallograft rejection and graft-v-host disease), and (11) other diseasesin which undesired inflammatory responses or immune disorders are to beinhibited, such as cardiovascular disease including atherosclerosis andrestenosis, myositis, neurodegenerative diseases (e.g., Alzheimer'sdisease), encephalitis, meningitis, hepatitis, nephritis, sepsis,sarcoidosis, allergic conjunctivitis, otitis, chronic obstructivepulmonary disease, sinusitis, Behcet's syndrome and gout and (12) immunemediated food allergies such as Celiac disease.

In another group of embodiments, diseases or conditions can be treatedwith modulators of CCR1 function. Examples of diseases to be treatedwith modulators of CCR1 function include cancers, cardiovasculardiseases, diseases in which angiogenesis or neovascularization play arole (neoplastic diseases, retinopathy and macular degeneration),infectious diseases (viral infections, e.g., HIV infection, andbacterial infections) and immunosuppressive diseases such as organtransplant conditions and skin transplant conditions. The term “organtransplant conditions” is meant to include bone marrow transplantconditions and solid organ (e.g., kidney, liver, lung, heart, pancreasor combination thereof) transplant conditions.

The compounds of the present invention are accordingly useful in theprevention and treatment of a wide variety of inflammatory andimmunoregulatory disorders and diseases.

Depending on the disease to be treated and the subject's condition, thecompounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by implantation (e.g., as when the compound is coupled to astent device), by inhalation spray, nasal, vaginal, rectal, sublingual,or topical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex and diet of the subject, as well as the mode and time ofadministration, rate of excretion, drug combination, and the severity ofthe particular condition for the subject undergoing therapy.

Diseases and conditions associated with inflammation, immune disorder,infection and cancer can be treated or prevented with the presentcompounds, compositions, and methods.

The compounds and compositions of the present invention can be combinedwith other compounds and compositions having related utilities toprevent and treat the condition or disease of interest, such asinflammatory or autoimmune disorders, conditions and diseases, includinginflammatory bowel disease, rheumatoid arthritis, osteoarthritis,psoriatic arthritis, polyarticular arthritis, multiple sclerosis,allergic diseases, psoriasis, atopic dermatitis and asthma, and thosepathologies noted above.

For example, in the treatment or prevention of inflammation orautoimmunity or for example arthritis associated bone loss, the presentcompounds and compositions may be used in conjunction with ananti-inflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non steroidal anti-inflammatory agent, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds and compositions may be administeredwith an analgesic listed above; a potentiator such as caffeine, an H2antagonist (e.g., ranitidine), simethicone, aluminum or magnesiumhydroxide; a decongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, ephinephrine, naphazoline,xylometazoline, propylhexedrine, or levo desoxy ephedrine; anantitussive such as codeine, hydrocodone, caramiphen, carbetapentane, ordextromethorphan; a diuretic; and a sedating or non sedatingantihistamine.

Likewise, compounds and compositions of the present invention may beused in combination with other drugs that are used in the treatment,prevention, suppression or amelioration of the diseases or conditionsfor which compounds and compositions of the present invention areuseful. Such other drugs may be administered, by a route and in anamount commonly used therefor, contemporaneously or sequentially with acompound or composition of the present invention. When a compound orcomposition of the present invention is used contemporaneously with oneor more other drugs, a pharmaceutical composition containing such otherdrugs in addition to the compound or composition of the presentinvention is preferred. Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients or therapeutic agents, in addition to a compound orcomposition of the present invention. Examples of other therapeuticagents that may be combined with a compound or composition of thepresent invention, either administered separately or in the samepharmaceutical compositions, include, but are not limited to: (a) VLA-4antagonists, (b) corticosteroids, such as beclomethasone,methylprednisolone, betamethasone, prednisone, prenisolone,dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone,salmeterol, salmeterol, salbutamol, formeterol; (c) immunosuppressantssuch as cyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolirnus(FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506type immunosuppressants, and mycophenolate, e.g., mycophenolate mofetil(CellCept®); (d) antihistamines (H1-histamine antagonists) such asbromopheniramine, chlorpheniramine, dexchloipheniramine, triprolidine,clemastine, diphenhydramine, diphenylpyraline, tripelennamine,hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine,cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non steroidal antiasthmatics (e.g., terbutaline, metaproterenol, fenoterol, isoetharine,albuterol, bitolterol and pirbuterol), theophylline, cromolyn sodium,atropine, ipratropium bromide, leukotriene antagonists (e.g.,zafmlukast, montelukast, pranlukast, iralukast, pobilukast andSKB-106,203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005);(f) non steroidal anti-inflammatory agents (NSAIDs) such as propionicacid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid,carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acidderivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin andzomepirac), fenamic acid derivatives (e.g., flufenamic acid,meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal),oxicams (e.g., isoxicam, piroxicam, sudoxicam and tenoxican),salicylates (e.g., acetyl salicylic acid and sulfasalazine) and thepyrazolones (e.g., apazone, bezpiperylon, feprazone, mofebutazone,oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2)inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h)inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compoundssuch as auranofin and aurothioglucose, (j) etanercept (Enbrel®), (k)antibody therapies such as orthoclone (OKT3), daclizumab (Zenapax®),basiliximab (Simulect®) and infliximab (Remicade®), (l) otherantagonists of the chemokine receptors, especially CCR5, CXCR2, CXCR3,CCR2, CCR3, CCR4, CCR7, CX₃CR1 and CXCR6; (m) lubricants or emollientssuch as petrolatum and lanolin, (n) keratolytic agents (e.g.,tazarotene), (o) vitamin D₃ derivatives, e.g., calcipotriene orcalcipotriol (Dovonex®), (p) PUVA, (q) anthralin (Drithrocreme®), (r)etretinate (Tegison®) and isotretinoin and (s) multiple sclerosistherapeutic agents such as interferon β-1β (Betaseron®), interferon(β-1α (Avonex®), azathioprine (Imurek®, Imuran®), glatiramer acetate(Capoxone®), a glucocorticoid (e.g., prednisolone) and cyclophosphamide(t) DMARDS such as methotrexate (u) other compounds such as5-aminosalicylic acid and prodrugs thereof; hydroxychloroquine;D-penicillamine; antimetabolites such as azathioprine, 6-mercaptopurineand methotrexate; DNA synthesis inhibitors such as hydroxyurea andmicrotubule disrupters such as colchicine. The weight ratio of thecompound of the present invention to the second active ingredient may bevaried and will depend upon the effective dose of each ingredient.Generally, an effective dose of each will be used. Thus, for example,when a compound of the present invention is combined with an NSAID theweight ratio of the compound of the present invention to the NSAID willgenerally range from about 1000:1 to about 1:1000, preferably about200:1 to about 1:200. Combinations of a compound of the presentinvention and other active ingredients will generally also be within theaforementioned range, but in each case, an effective dose of each activeingredient should be used.

VI. Examples

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR spectra wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In the examples, a single m/e value isreported for the M+H (or, as noted, M−H) ion containing the most commonatomic isotopes. Isotope patterns correspond to the expected formula inall cases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP1100 HPLC for sample delivery. Normally the analyte was dissolvedin methanol at 0.1 mg/mL and 1 microliter was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. A sample of useful routes to the azaindazole derivatives andcertain compounds of the invention are provided below or elsewherewithin the present application. In the descriptions of the synthesesthat follow, some of the arylpiperazine and heteroaromatic subunitprecursors were obtained from commercial sources. These commercialsources include Aldrich Chemical Co., Acros Organics, Ryan ScientificIncorporated, Oakwood Products Incorporated, Lancaster Chemicals, SigmaChemical Co., Lancaster Chemical Co., TCI-America, Alfa Aesar, DavosChemicals, and GFS Chemicals. Certain relevant arylpiperazine compoundscan be commercially obtained. Others could be prepared as described inU.S. patent application Ser. No. 11/008,774, the contents of which ishereby incorporated in its entirety for all purposes. Also, standardchemistries have been employed to link the arylpiperazine andheteroaromatic subunits (whether commercially obtained or prepared bythe methods below) using a suitably optimized linker, such as the acetylunit described in the body of this invention.

One skilled in the art will also recognize that alternative methods maybe employed to synthesize the target compounds of this invention, andthat the approaches described within the body of this document are notexhaustive, but do provide broadly applicable and practical routes tocompounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

Regioisomerism is a common property in organic chemistry, and isespecially common with regards to certain structural types providedherein. Those skilled in the art will recognize, with respect to thecompounds described herein, that the coupling reactions with theheteroaromatic ring systems can lead to either one of or a mixture ofdetectable regioisomers.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

Example 1 Synthesis of 1H-pyrazolo[3,4-b]pyridine

2-Chloro-3-formylpyridine (15.02 g, 106 mmol, 1 equiv), hydrazine (10mL, excess), and dioxane (90 mL) were combined in a sealed tube andheated at 150° C. for 16 hr. After cooling to room temperature, thesolvent was evaporated in vacuo to provide a crude residue which wasdiluted with dichloromethane (600 mL). The organic solution was washedwith water (50 mL), brine (50 mL) and dried over anhydrous sodiumsulfate. The solvent was removed in vacuo to provide1H-pyrazolo[3,4-b]pyridine as a yellow powder which was used withoutfurther purification: LCMS (ES) M+H 120.3, R_(f) 0.20 min (AgilentZorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 mingradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formicacid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 2 Synthesis of 3-Thiazol-2-yl-1H-pyrazolo[3,4-b]pyridine

To a suspension of 2-chloro-3-[(2-thiazolyl)carbonyl]pyridine (257.5 mg,1.2 mmol, 1 equiv) in dioxane (3 mL) in a sealed tube was addedhydrazine (2 mL). The mixture was heated at 150° C. overnight, cooled toroom temperature and concentrated in vacuo to provide a crude residue.The resultant residue was diluted with dichloromethane (300 mL), washedwith water (50 mL) and brine (50 mL). The organic layer was separated,dried over sodium sulfate, filtered and concentrated in vacuo to provide3-thiazol-2-yl-1H-pyrazolo[3,4-b]pyridine (212.3 mg) as a yellow powderwhich used without further purification: LCMS (ES) M+H 203.5, R_(f) 2.68min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a2.5 min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile).

Example 3 Synthesis of 1H-Pyrazolo[3,4-b]pyridin-3-ylamine

2-Chloro-3-cyanopyridine (2.77 g), hydrazine (5 mL), and dioxane (100mL) were combined in a sealed tube and heated at 150° C. for 16 hr. Thereaction mixture was cooled to room temperature and concentrated invacuo to provide a crude residue. The resultant residue was dissolved inethyl acetate (100 mL) and washed with saturated NaCl solution (50 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford 1H-pyrazolo[3,4-b]pyridin-3-ylamine as ayellow solid which was used without further purification.

Example 4 Synthesis of 1H-pyrazolo[3,4-c]pyridine

Preparation of 3-N-acetylamino-4-methylpyridine: To solution of3-amino-4-methylpyridine (540.2 mg, 5.0 mmol, 1 equiv) indichloromethane (20 mL) was added pyridine (0.8 mL, 10.0 mmol, 2 equiv)and acetic anhydride (0.57 mL, 6.0 mmol, 1.2 equiv). The resultantsolution was stirred at room temperature for 16 h and concentrated invacuo to provide a crude residue. The residue was diluted withdichloromethane (200 mL), and washed with saturated sodium bicarbonateaqueous solution (50 mL) and brine (50 mL). The organic layer wasseparated, dried over sodium sulfate, and concentrated in vacuo to yield3-acetylamino-4-methylpyridine (400.2 mg) as yellow solid which was usedwithout further purification.

Preparation of 1-pyrazolo[3,4-c]pyridin-1-yl-ethanone: To a suspensionof 3-acetylamino-4-methylpyridine (301.5 mg, 2.0 mmol, 1 equiv) intoluene (3 mL) was added tert-butyl nitrite (t-BuONO) (420 μL, 3.2 mmol,1.6 equiv), acetic anhydride (560 μL, 6.0 mmol, 3 equiv) and potassiumacetate (235.2 mg, 2.4 mmol, 1.2 equiv). The resultant mixture washeated at 80° C. for 2 hours, cooled to room temperature, and dilutedwith ethyl acetate (200 mL). The mixture was washed with saturatedsodium bicarbonate solution (50 mL), water (50 mL) and brine (50 mL).The organic layer was dried over sodium sulfate and concentrated invacuo to provide a crude residue. The residue was purified by flashchromatography (silica, 15% ethyl acetate/hexane to 50% ethylacetate/hexane) to give 1-pyrazolo[3,4-c]pyridin-1-yl-ethanone (20.2 mg)which was used without further purification.

Synthesis of 1H-pyrazolo[3,4-c]pyridine: To a solution of1-pyrazolo[3,4-c]pyridin-1-yl-ethanone (20.2 mg, 0.17 mmol, 1 equiv) intetrahydrofuran (2 mL) and methanol (0.5 mL) was added sodium hydroxideaqueous solution (2M, 0.25 mL). The reaction solution was stirred atroom temperature for 1 hr and then concentrated in vacuo to provide acrude residue. The crude residue was diluted with water (20 mL) andextracted with dichloromethane (2×100 mL). The combined organic layerswere washed with brine, dried over sodium sulfate, and concentrated invacuo to provide 1H-pyrazolo[3,4-c]pyridine as white powder, which usedwithout further purification: LCMS (ES) M+H 120.3, R_(f) 0.22 min(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile).

Example 5 Synthesis of 3-Iodo-1H-pyrazolo[3,4-b]pyridine

To a solution of 1H-pyrazolo[3,4-b]pyridine (500.0 mg, 4.2 mmol, 1equiv) in DMF (10 mL) at 0° C., was added iodine (2.13 g, 8.4 mmol, 2equiv) and potassium hydroxide (943 mg, 16.8 mmol, 4 equiv). Theresultant mixture was allowed to warm to room temperature and stirredfor 1 hour. The reaction solution was slowly quenched with saturatedsodium thiosulfate (Na₂S₂O₅) solution (10 mL), and extracted with ethylacetate (2×200 mL). The combined organic layers were washed with water(3×50 mL), brined (50 mL), dried over sodium sulfate and concentrated invacuo to give 3-Iodo-1H-pyrazolo[3,4-b]pyridine (1.02 g) as a yellowpowder which was used without further purification: LCMS (ES) M+H 246.2,R_(f) 2.17 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/minflow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash at100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile).

Example 6 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-1-ylethanoneand1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-2-ylethanone

2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]ethanone(arylpiperazine) (4.81 g, 14.32 mmol, 1 equiv),1H-pyrazolo[3,4-b]pyridine (2.27 g, 17.18 mmol, 1.2 equiv), andpotassium carbonate (20.00 g, 143.2 mmol, 10 equiv) were dissolved indimethylformamide (DMF) (10 mL) and heated at 80° C. for 1 hour, thencooled to room temperature. The resultant mixture was diluted with ethylacetate (300 mL), and washed with water (3×150 mL) and brine (100 mL).The organic layer was dried (Na₂SO₄) and concentrated in vacuo toprovide a crude residue. The crude residue was purified by flashchromatography (silica, 100% ethyl acetate with 1% triethylamine to 100%acetone with 1% triethylamine) provided1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-1-ylethanone(2.3 g) and1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-2-ylethanone(2.5 g).

For1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-1-yl-ethanone;LCMS (ES) M+H 418.5, R_(f) 2.34 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile): For1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-2-ylethanone;LCMS (ES) M+H 418.5, R_(f) 2.00 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 7 Synthesis of1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

Preparation of1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:This compound was synthesized according to the synthetic procedureoutlined in Example 6.

Synthesis of1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:To a solution of oxazole (40 μL, 0.54 mmol, 3 equiv) in tetrahydrofuran(1 mL) under nitrogen atmosphere, was added dropwise n-butyl lithium(2.5 M in Hexane, 220 μL, 0.54 mmol, 3 equiv.). The resultant mixturewas stirred at −78° C. for an additional 30 min followed by the additionof ZnCl₂ (0.5M in THF, 1.5 mL, 0.72 mmol, 4 equiv.). The reactionsolution was allowed to warm to 0° C. and stirred 1 hr followed by theaddition of1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(100.2 mg, 0.18 mmol, 1 equiv) and palladiumtetrakis(triphenylphosphine) (22.3 mg, 0.018, 0.1 equiv). The reactionmixture was then heated to reflux for 48 hr, cooled to room temperatureand diluted with ethyl acetate (150 mL). The reaction mixture was washedwith water (20 mL), brine (20 mL), dried over sodium sulfate, andconcentrated in vacuo to provide the crude product. Purification bypreparative HPLC provided the desired product1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanoneas a white powder (38.5 mg): LCMS (ES) M+H 485.5, R_(f) 2.56 min(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile).

Example 8 Synthesis of1-[4-(4-chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-1-ylethanoneand1-[4-(4-chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-2-ylethanone

The two title compounds were synthesized according to the syntheticprocedure as outlined in Example 6: For1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-1-ylethanone;¹H NMR (400 MHz, CDCl₃) δ 8.57 (dd, 1H), 8.11 (s, 1H), 8.09 (dd, 1H),7.22 (d, 1H), 7.17 (dd, 1H), 6.49 (d, 1H), 6.42 (dd, 1H), 5.44 (s, 2H),3.92 (s, 3H), 3.79 (m, 4H), 3.18 (m, 4H); MS (M+H)⁺: 386.5: For1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine-2-ylethanone;¹H NMR (400 MHz, CDCl₃) δ 8.65 (dd, 1H), 8.12 (s, 1H), 8.02 (dd, 1H),7.20 (d, 1H), 7.03 (dd, 1H), 6.45 (d, 1H), 6.40 (dd, 1H), 5.35 (s, 2H),3.88 (s, 3H), 3.87 (m, 2H), 3.79 (m, 2H), 3.15 (m, 4H); MS (M+H)⁺:386.5.

Example 9 Synthesis of 1H-Pyrazolo[4,3-c]pyridine

1H-Pyrazolo[4,3-c]pyridine was prepared according to the procedureoutlined in Example 1.

Example 10 Synthesis of1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[4,3-c]pyridine-1-yl-ethanoneand1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[4,3-c]pyridine-2-yl-ethanone

The two title compounds were synthesized according to the procedure asoutlined in Example 6. For1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[4,3-c]pyridine-1-yl-ethanone:LCMS (ES) M+H 418.5, R_(f) 1.74 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile): For1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-pyrazolo[4,3-c]pyridine-2-yl-ethanone;LCMS (ES) M+H 418.5, R_(f) 1.69 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 11 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridine-1-yl-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridine-2-yl-ethanone

The two title compounds were synthesized according to the procedureoutlined in Example 6: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridine-1-yl-ethanone;¹H NMR (400 MHz, CDCl₃) 9.02 (s, 1H), 8.34 (d, 1H), 8.09 (d, 1H), 7.63(dd, 1H), 7.22 (d, 1H), 6.48 (d, 1H), 6.42 (dd, 1H), 5.38 (s, 2H), 3.88(s, 3H), 3.79 (m, 4H), 3.14 (m, 4H) MS (M+H)⁺, 386.5: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridine-2-yl-ethanone;¹H NMR (400 MHz, CDCl₃)) 9.22 (s, 1H), 8.13 (d, 1H), 8.10 (d, 1H), 7.50(dd, 1H), 7.19 (d, 1H), 6.45 (d, 1H), 6.39 (dd, 1H), 5.37 (s, 2H), 3.85(s, 3H), 3.76 (m, 4H), 3.14 (m, 4H). MS (M+H)⁺, 386.5.

Example 12 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanoneand1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone

The two title compounds were synthesized according to the procedureoutlined in Example 6. For1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone:LCMS (ES) M+H, 404.5, R_(f) 2.14 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile): For1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone;LCMS (ES) M+H, 404.5, R_(f) 1.76 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 13 Synthesis of1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 501.5, R_(f) 2.82 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 14 Synthesis 3-pyrid-2-yl-1H-pyrazolo[3,4-b]pyridine

The title compound was synthesized according to the procedure outlinedin Example 2.

Example 15 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 495.54, R_(f) 2.73 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 16 Synthesis of 3-Chloro-1H-pyrazolo[3,4-b]pyridine

1H-pyrazolo[3,4-b]pyridine (89 mg) and N-chlorosuccinimide (220 mg) werecombined in CH₂Cl₂ (4 mL) and heated at 45° C. for 16 hr, then cooled toroom temperature. The resultant mixture was purified by flashchromatography (silica gel, 50% hexane/ethyl acetate) to afford3-chloro-1H-pyrazolo[3,4-b]pyridine.

Example 17 Synthesis of1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-chloropyrazolo[3,4-b]pyridin-1-yl)ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 420.5, R_(f) 2.37 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 18 Synthesis of 3-Methyl-1H-pyrazolo[3,4-b]pyridine

To a solution of 2-chloro-3-cyanopyridine (139 mg) in tetrahydrofuran (5mL) at 0° C. was added dropwise a solution of MeMgBr (3M in ether, 0.67mL). The resultant mixture was warmed to room temperature and stirredfor 3 hr. The reaction solution was cooled to 0° C. and to it was addedaqueous HCl solution (2M, 5 mL). The reaction solution was then stirredan additional 16 hr at room temperature and then neutralized by theaddition of saturated sodium bicarbonate (NaHCO₃) solution. The reactionsolution was filtered to remove any precipitates and the filtrate waswashed with ethyl acetate (3×10 mL) and aqueous brine (NaCl) solution(10 mL). The organic layer was dried over sodium sulfate (Na₂SO₄),filtered and concentrated in vacuo to give 3-acetyl-2-chloropyridine asa yellow powder which was used without further purification. The titlecompound (3-Methyl-1H-pyrazolo[3,4-b]pyridine) was synthesized from3-acetyl-2-chloropyridine according to the procedure outlined in Example2.

Example 19 Synthesis of1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-methylpyrazolo[3,4-b]pyridin-1-yl)ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 400.5, R_(f) 2.12 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; 0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 20 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-(3-methylpyrazolo[3,4-b]pyridin-1-yl)ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 432.5, R_(f) 2.42 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 21 Synthesis of1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 463.5, R_(f) 2.32 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 22 Synthesis of1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)ethanone

The title compound was synthesized according the procedure outlined inExample 6: LCMS (ES) M+H 469.5, R_(f) 2.43 min (Agilent Zorbax SB-C18,2.1 50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 23 Synthesis of2-(3-Aminopyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]ethanone

1H-Pyrazolo[3,4-b]pyridin-3-ylamine (67 mg),2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-ethanone(167 mg) and K₂CO₃ (414 mg) were combined in DMF (1 mL) and heated at80° C. for 2 hr, then cooled to room temperature. The resultant mixturewas purified by preparative HPLC to provide2-(3-Aminopyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]ethanoneas a yellow powder. LCMS (ES) M+H 433.5, R_(f) 2.06 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 24 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

The title was synthesized according to the procedure outlined in Example7: LCMS (ES) M+H 453.5, R_(f) 2.20 min (Agilent Zorbax SB-C18, 2.1×50mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% Bwith a 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 25 Synthesis of 3-Fluoro-1H-pyrazolo[3,4-b]pyridine

The title was synthesized according to the procedure outlined in Example5 using SelectFluor™(1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate)) as the electrophile.

Example 26 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-fluoro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

The title compound was synthesized according to the procedure outlinedin Example 6: LCMS (ES) M+H 404.5, R_(f) 2.27 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.08% formic acid/5% water/94.9%acetonitrile).

Example 27 Synthesis of1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile

1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(128 mg) and CuCN (112 mg) were combined in N-methylpyridone (NMP) (1mL) and heated at 165° C. for 16 hr, then cooled to room temperature.The reaction mixture was purified on preparative HPLC to afford1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrileas a white powder: LCMS (ES) M+H 411.5, R_(f) 2.33 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 mL/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 28 Synthesis of 1H-pyrazolo[4,3-b]pyridine

1H-pyrazolo[4,3-b]pyridine was synthesized according to the procedureoutlined in Example 4: LCMS (ES) M+H 120.3.

Example 29 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-1-yl-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-2-yl-ethanone

The two title compounds were synthesized according to the procedureoutlined in Example 6: For1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-1-yl-ethanone;¹H NMR (400 MHz, DMSO-d₆) δ 8.71 (dd, 1H), 8.26 (s, 1H), 8.04 (dd, 1H),7.36 (dd, 1H), 7.20 (d, 1H), 6.69 (d, 1H), 6.51 (dd, 1H), 5.57 (s, 2H),3.82 (s, 3H), 3.73 (m, 2H), 3.59 (m, 2H), 3.31 (m, 2H), 3.19 (m, 2H).LCMS (ES) M+H 386.5, R_(f) 1.84 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile). For1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-2-yl-ethanone;¹H NMR (400 MHz, CDCl₃) δ 8.55 (d, 1H), 8.34 (s, 1H), 7.99 (d, 1H), 7.19(m, 2H), 6.44 (d, 1H), 6.40 (dd, 1H), 5.34 (s, 2H), 3.86 (s, 3H), 3.77(m, 2H), 3.72 (m, 2H), 3.13 (m, 4H). LCMS (ES) M+H 386.5, R_(f) 1.69 min(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile).

Example 30 Synthesis of 2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-aceticacid

Preparation of pyrazolo[3,4-b]pyridin-2-yl-acetic acid ethyl ester: Thiscompound was synthesized according to the procedure outlined in Example6, using chloro-acetic acid ethyl ester in place of2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]ethanone.

Preparation of (3-Chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic acid ethylester: To a solution of pyrazolo[3,4-b]pyridin-2-yl-acetic acid ethylester 57 (40.2 mg, 0.2 mmol, 1 equiv) in 1 mL of dichloromethane wasadded NCS (32.7 mg, 1.2 mmol, 1.2 equiv). The resultant mixture washeated at 70° C. for 30 min., cooled to room temperature, and dilutedwith 100 mL of dichloromethane. The organic solution was washed with 50mL of saturated sodium bicarbonate aqueous solution, and 50 mL of brine.The organic layer was separated and dried over sodium sulfate.Evaporation of solvent in vacuo gave 46.7 mg of(3-chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic acid ethyl ester as yellowsolid.

Synthesis of 2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid:(3-Chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic acid ethyl ester wastreated with 1N lithium hydroxide (LiOH) (1 equiv) in 1 mL of MeOH toprovide 2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid, which wasused as directly in subsequent reactions without further purification:LCMS (ES) M+H 212.0, R_(f) 0.34 min (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B witha 1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 31 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4-b]pyridine-2-yl)-ethanone

The title compound was synthesized according to standard amide formationconditions using2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HATU) as the coupling reagent: LCMS (ES) M+H 420.4,R_(f) 2.17 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/minflow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash at100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile.

Example 32 Synthesis of 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-aceticacid

Preparation of 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethylester: To a solution of pyrazolo[3,4-b]pyridin-1-yl-acetic acid ethylester (205.4 mg, 1 mmol, 1 equiv) in 10 mL of dichloromethane at 0° C.,was added meta-chloroperoxybenzoic acid (mCPBA) (345.3 mg, 1.5 mmol, 1.5equiv). The resultant mixture was allowed to warm to room temperature,and the reaction was stirred overnight. 1 mL of pyridine was added tothe reaction mixture, and the mixture was stirred for another 30 minbefore the solvent was removed to provide a residue. The residue wasdiluted with 200 mL of dichloromethane, and washed with 1 N NaOH aqueoussolution (10 mL×2), brine (20 mL). The organic layer was separated anddried over sodium sulfate. Evaporation in vacuo gave2-(pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethyl ester as paleyellow solid, which was used without further purification: LCMS (ES) M+H222.4, R_(f) 1.48 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 minwash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%formic acid/5% water/94.9% acetonitrile.

Synthesis of 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid:2-(pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethyl ester wastreated with 1N LiOH (1 equiv) in 1 mL of methanol (MeOH) to provide2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid: LCMS (ES) M+H194.2, R_(f) 0.22 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 minwash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%formic acid/5% water/94.9% acetonitrile

Example 33 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)piperazin-1-yl]-2-(pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-ethanone

The title compound was prepared according to standard amide formationconditions as described in Example 43 using HATU as the couplingreagent: LCMS (ES) M+H 402.5, R_(f) 1.54 min (Agilent Zorbax SB-C18,2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of 20% to100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile.

Example 34 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

2-chloro-3-cyano-6-picoline was reduced by diisobutylaluminum hydride(DIBAL-H) following a literature procedure (Baker et. al., J. Org.Chem., 1980, 45, 1354-1362.) followed by the hydrazine condensationprotocol as described in Example 1 to provide the corresponding6-Methyl-1H-pyrazolo[3,4-b]pyridine, which was then subjected to thealkylation protocol in described in Example 6 to provide title compoundas a white powder: LCMS (ES) M+H 400.5, R_(f) 2.161 min (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 min gradient of20% to 100% B with a 1.1 min wash at 100% B, A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile).

Example 35 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-ethanone

Preparation of (6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid ethylester: To a solution of 1H-6-methyl-pyrazolo[3,4-b]pyridine (1 mmol, 1eq.) in 3 mL of THF was added NaH (1.5 mmol, 1.5 eq.) portion by portionat 0° C. under nitrogen. The resultant mixture was stirred at 0° C. for10 minutes followed by the slow addition of 2-chloro ethyl acetate(excess) at 0° C. The resultant mixture was slowly to warmed to rt, andstirred for another 2 h. To the reaction mixture was added saturatedNH₄Cl aq. solution, and aqueous mixture was extracted with 300 mL ofEtOAc. The organic extract was separated and washed with sat. sodiumbicarbonate aq. solution, brine solution, filtered and dried over sodiumsulfate. The organic solvent was removed in vacuo, and the crude residuewas purified by silica gel chromatography to provide 50.2 mg desiredproduct: HPLC retention time=0.78 minutes (Agilent Zorbax SB-C18, 2.1×50mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20%to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=220.1. found=220.4.

Preparation of (6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid: Thiscompound was synthesized according to standard ester hydrolysis protocolas described in Example 30 using 1N LiOH as the base. The isolatedproduct was used in the next step without purification.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-ethanone:The title compound was synthesized according to standard peptidecoupling protocol using HATU as the coupling reagent: ¹H NMR (400 MHz,CDCl₃) δ 8.04 (s, 1H), 7.96 (d, 1H), 7.20 (d, 1H), 6.95 (d, 1H), 6.45(d, 1H), 6.40 (dd, 1H), 5.29 (s, 2H), 3.92 (m, 2H), 3.88 (s, 3H), 3.78(m, 2H), 3.13 (m, 4H). LCMS observed for (M+H)⁺: 400.5.

Example 36 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-morpholin-4-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

A mixture of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(102.4 mg), morpholine (0.20 mL), Xantphos (35 mg), Pd₂(dba)₃ (18.3 mg)and Cs₂CO₃ (97 mg) in THF (1 mL) was heated to 80° C. for 12 h. Thereaction mixture was allowed to cool to room temperature, diluted byEtOAc (3 mL) and filtered. The filtrate was evaporated in vacuo. Thecrude residue was purified by flash chromatography (silica,Hexane/EtOAc) to provide the title compound as a white powder: LCMS (ES)M+H 471.6, R_(f) 2.043 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35°C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1min wash at 100% B, A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile).

Example 37 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-1-yl-ethanone

The title compound was synthesized following the alkylation protocol asdescribed in Example 6: LCMS (ES) M+H 418.4, R_(f) 2.055 min (AgilentZorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1 ml/min flow rate, a 2.5 mingradient of 20% to 100% B with a 1.1 min wash at 100% B, A=0.1% formicacid/5% acetonitrile/94.9% water, B=0.08% formic acid/5% water/94.9%acetonitrile).

Example 38 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-oxy-pyrazolo[3,4-c]pyridin-1-yl)-ethanone

A mixture of 6-azaindazole (119 mg), H₂O₂ (0.2 mL) in acetic acid (5 mL)was heated to 60° C. for 2 h. The resultant mixture was cooled to roomtemperature and concentrated in vacuo. The crude residue was dissolvedin EtOAc (10 mL), washed with sat. aqueous NaHCO₃ solution (3 mL), dried(Na₂SO₄), filtered and evaporated in vacuo. The crude product (theN-oxide) was subjected to the alkylation protocol as described inExample 6 to provide the title compound as a white powder: LCMS (ES) M+H402.4, R_(f) 2.147 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 minwash at 100% B, A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%formic acid/5% water/94.9% acetonitrile).

Example 39 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[4,3-c]pyridine-2-yl)-ethanone

Preparation of 1H-4-chloro-pyrazolo[4,3-c]pyridine: To a mixture of2-chloro-4-iodopyridine-3-carbaldehyde (6.24 mmol, 1 eq.) and 5 mL ofethanol was added 4 mL of hydrazine (excess), the resultant mixture wasstirred at rt for 6 h. The reaction solution was concentrated in vacuo,and the crude residue was diluted with 50 mL of water, and extractedwith 500 mL of dichloromethane. The organic layer was then washed withbrine, dried over anhydrous sodium sulfate, filtered and concentrated toprovide a crude residue. To the crude residue was dissolved with 10 mLof dichloromethane and stirred for 5 minutes. The precipitated solidswere isolated by filtration, washed with 2 mL of dichloromethane, anddried in vacuo to provided 350.2 mg of1H-4-chloro-pyrazolo[4,3,c]pyridine: HPLC retention time=0.44 minutes(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate,a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100% B(A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile); MS (ES) M+H expected=154.0. found=154.3.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[4,3-c]pyridine-2-yl)-ethanone:Using 1H-4-chloro-pyrazolo[4,3-c]pyridine, the title compound wassynthesized according to alkylation protocol in Example 6: ¹H NMR (400MHz, CDCl₃) δ 8.18 (d, 1H), 7.31 (dd, 1H), 7.21 (d, 1H), 6.47 (d, 1H),6.42 (dd, 1H), 5.28 (s, 2H), 3.88 (s, 3H), 3.77 (m, 4H), 3.14 (m, 4H).LCMS observed for (M+H)⁺: 420.4.

Example 40 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-2-yl)-ethanone

Preparation of 1H-4-iodo-pyrazolo[3,4,b]pyridine: To a mixture of2-chloro-4-iodopyridine-3-carbaldehyde (6.24 mmol, 1 eq.) and 5 mL ofethanol was added 4 mL of hydrazine (excess), the resultant mixture wasstirred at rt for 6 h. The reaction mixture was concentrated in vacuoand crude residue was diluted with 50 mL of water, and extracted with500 mL of dichloromethane. The organic layer was washed with brine,dried over anhydrous sodium sulfate and concentration in vacuo toprovide a crude residue. To this residue was added 10 mL ofdichloromethane, the resultant mixture was stirred for 5 minutes whichresulted in the precipation of the undesired cyclization isomer(1H-4-chloro-pyrazolo[4,3,c]pyridine) which was removed by filtration.The filtrate was concentrated in vacuo, and purified be by silica gelcolumn (35% acetone in hexane to 50% acetone in hexane) to provide 250.0mg of 1H-4-iodo-pyrazolo[3,4,b]pyridine with a purity around 85%, whichwas used without further purification: HPLC retention time=1.22 minutes(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate,a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100% B(A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile); MS (ES) M+H expect=246.0. found=246.1.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-2-yl)-ethanone:The title compounds were synthesized according to the standardalkylation procedure described in Example 6. For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-1-yl)-ethanone:HPLC retention time=2.50 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=512.0. found=512.4; For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-2-yl)-ethanone:HPLC retention time=2.23 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=512.0. found=512.4

Example 41 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methylsulfonyl-pyrazolo[4,3-c]pyridine-1-yl)-ethanone

A mixture of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c]pyridine-1-yl)-ethanone(0.1 mmol, 1 eq.), CuI (0.3 mmol, 3 eq.) and NaSO₂Me (0.3 mmol, 1 eq.)in 1 mL of DMSO was heated at 80° C. for 2 h. The reaction solution wascooled to rt, and diluted with 20 mL of sat. NH₄Cl aq. solution and 200mL of EtOAc. The diluted mixture was stirred vigorously for 2 h. Theorganic layer was then separated, washed with brine, dried over sodiumsulfate, filtered and concentrated in vacuo to provide the crudeproduct. Purification by HPLC provided 40.2 mg desired product: ¹H NMR(400 MHz, CDCl₃) δ 8.76 (d, 1H), 8.49 (d, 1H), 7.65 (d, 1H), 7.23 (d,1H), 6.50 (d, 1H), 6.44 (dd, 1H), 5.53 (s, 2H), 3.89 (s, 3H), 3.79 (m,4H), 3.22 (m+s, 4H+3H). LCMS observed for (M+H)⁺: 464.4.

Example 42 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-amidomethyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

Preparation of (3-methyl-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethylester: This compound was synthesized following the alkylation protocolsimilar to the one described in Example 6: HPLC retention time=2.06minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/minflow rate, a 2.5 minute gradient of 0% to 100% B with a 1.1 minute washat 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile); MS (ES) M+H expect=220.1.found=220.4.

Preparation of [3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic acidethyl ester: A mixture of (3-methyl-pyrazolo[3,4-b]pyridine-1-yl)ethylacetate (2.5 mmol, 1 eq.), NBS (3.0 mmol, 1.2 eq.), and benzoyl peroxide(0.05 mmol, 0.02 eq.) in 10 mL of CCl₄ was refluxed for 1.5 h. Theresultant mixture was cooled to rt, and diluted with 500 mL of EtOAc.The resultant solution was then washed with 100 mL of sat. sodiumbicarbonate aqueous solution, brine solution, dried over anhydroussodium sulfate, filtered and concentrated in vacuo. The crude residuewas purified by silica gel chromatography (20% EtOAc in hexane to 35%EtOAc in hexane) to provide 450.2 mg of the desired product: HPLCretention time=2.50 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35°C.) using 1 ml/min flow rate, a 2.5 minute gradient of 0% to 100% B witha 1.1 minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=298.0. found=298.3.

Preparation of (3-azido-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethylester: A mixture of (3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)aceticacid ethyl ester (0.5 mmol, 1 eq.) and sodium azide (1 mmol, 2 eq.) in 1mL of DMF was heated at 80° C. for 1 h. The resultant mixture was cooledto rt, diluted with 150 mL of EtOAc, washed with water (40 mL×3), brine,dried over anhydrous sodium sulfate, filtered and concentrated in vacuo.The solvent was removed in vacuo to provide 135.2 mg desired product:HPLC retention time=1.84 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=261.1. found=261.4.

Preparation of [3-(azidomethyl)-pyrazolo[3,4-b]pyridine-1-yl)aceticacid: This compound was synthesized according to standard hydrolysisprotocol as described in Example 30 using 1 N LiOH: HPLC retentiontime=1.94 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 0% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=233.1. found=233.4.

Preparation of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-azidomethyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone:This compound was synthesized according to standard peptide couplingprocedure as described below in Example 43 using HATU as the couplingreagent: HPLC retention time=2.36 minutes (Agilent Zorbax SB-C18, 2.1×50mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20%to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=441.2. found=441.5.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-amidomethyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone:To a solution of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-azidomethyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone(0.21 mmol, 1 eq.) in 2 mL of THF was added dropwise at rt a solution oftris(2-carboxyethyl)phosphine HCl salt in 0.5 mL of water. The resultantmixture was stirred at rt for 30 min. The reaction solution wasconcentrated in vacuo, and the crude residue was diluted with 150 mL ofdichloromethane, washed with 25 mL of water, brine, and dried oversodium sulfate. The solvent was removed in vacuo, and the crude residuewas purified by HPLC to provide 26.2 mg final product: ¹H NMR (400 MHz,CDCl₃) δ 8.17 (dd, 1H), 8.15 (dd, 1H), 7.22 (d, 1H), 7.12 (dd, 1H), 6.50(d, 1H), 6.44 (dd, 1H), 5.40 (s, 2H), 4.25 (s, 2H), 3.89 (s, 3H), 3.77(m, 4H), 3.19 (m, 4H). LCMS observed for (M+H)⁺: 416.4.

Example 43 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonicacid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

Preparation of [3-(sulfonicacid-methyl)-pyrazolo[3,4-b]pyridine-1-yl]acetic acid ethyl ester: Amixture of (3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic acidethyl ester (0.13 mmol, 1 eq.) and sodium sulfite (1.8 mmol, excess) ina mixture of 1 mL of DMF and 0.5 mL of water was heated at 80° C. forone hour. The resultant solution was cooled to rt, and the solvent wasremoved in vacuo. The residue was extracted with 1:1 MeOH:CH₂Cl₂ (30mL×3). The combined organic extracts were dried in vacuo, and the cruderesidue was used without further purification: HPLC retention time=1.63minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/minflow rate, a 2.5 minute gradient of 0% to 100% B with a 1.1 minute washat 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile); MS (ES) M+H expect=300.1.found=300.5.

Preparation of [3-(sulfonicacid-methyl)-pyrazolo[3,4-b]pyridine-1-yl]acetic acid: This compound wassynthesized according to standard hydrolysis protocol as described inExample 30 using 1 N LiOH as the base. The crude product was usedwithout further purification.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonicacid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone: A mixture ofsulfonic acid (100.2 mg, contain lots of inorganic salt),1H-4-(4-chloro-3-methoxy-phenyl)piperazine 2× HCl salt (0.37 mmol,excess), and HATU (0.37 excess) was suspended in 3 mL of pyridine,stirred at rt for 3 h. The pyridine solvent was removed in vacuo, andthe crude residue was extracted with dichloromethane (10 mL×3). Theorganic extracts were removed, and the crude residue was purified byHPLC to provide 10 mg of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonicacid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone: HPLC retentiontime=0.28 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=480.1. found=480.5.

Example 44 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

Preparation of (3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)acetic acidethyl ester: To a solution of(3-iodo-5-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethyl ester (0.61, 1eq.) in 2 mL of DMF was added N-chlorosuccinimide (NCS) (0.73, 1.2 eq.)as a solid. The resultant mixture was heated at 70° C. for 3 h. Thereaction mixture was cooled to rt, and diluted with 250 mL of EtOAc. Thediluted mixture was then washed with water (100 mL×3), brine, dried oversodium sulfate, filtered and concentrated in vacuo. The crude residuewas purified by silica gel chromatography (15% EtOAc to 75% EtOAc inhexane) to provide 100.4 mg white solid as final product: HPLC retentiontime=2.48 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=365.9. found=366.3.

Preparation of (3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)aceticacid: This compound was synthesized according to standard hydrolysisprocedure as described in Example 30 using 1N LiOH: HPLC retentiontime=1.78 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=337.9. found=337.9.

Preparation of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanone:The title compound was synthesized according to standard peptidecoupling protocol as described in Example 43 using HATU as the couplingreagent: HPLC retention time=2.71 minutes (Agilent Zorbax SB-C18, 2.1×50mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20%to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=546.0. found=546.4.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanone:To a solution of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanone(0.037 mmol, 1 eq.) in 1.5 mL of dichloromethane under a nitrogenatmosphere cooled to −40° C., was added dropwise, 30 μl of 2.0 Msolution of isopropyl magnesium chloride (0.056 mmol, 1.5 eq.) in THF.The resultant mixture was for 30 minutes at −40° C. followed by dropwiseaddition of an ammonium chloride aqueous (aq) solution at lowtemperature. The reaction solution was warmed to rt, diluted with 200 mLof EtOAc, washed with 50 mL of water, brine, dried over sodium sulfate,filtered and concentrated in vacuo. The crude residue was purified byHPLC to provide 5 mg final product: ¹H NMR (400 MHz, CDCl₃) δ 8.44 (d,1H), 8.05 (m, 1H), 7.23 (d, 1H), 6.50 (d, 1H), 6.43 (dd, 1H), 5.42 (s,2H), 3.89 (s, 3H), 3.76 (m, 4H), 3.20 (m, 4H). LCMS observed for (M+H)⁺:421.1.

Example 45 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-2-yl)-ethanone

Preparation of 1H-4-chloro-pyrazolo[3,4-d]pyrimidine: This compound wassynthesized according to standard hydrazine cyclization protocol asdescribed in Example 1: HPLC retention time=0.36 minutes (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minutegradient of 0% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formicacid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=155.0. found=155.0.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-2-yl)-ethanone:These compounds were synthesized using1H-4-chloro-pyrazolo[3,4-d]pyrimidine following the alkylation procedureas described in Example 6: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-1-yl)-ethanone;¹H NMR (400 MHz, CDCl₃) δ 8.76 (s, 1H), 8.22 (s, 1H), 7.22 (d, 1H), 6.54(d, 1H), 6.44 (dd, 1H), 5.41 (s, 2H), 3.88 (s, 3H), 3.77 (m, 4H), 3.23(m, 4H), LCMS observed for (M+H)⁺: 421.1: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-c]pyrimidine-2-yl)-ethanone;HPLC retention time=1.70 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=421.1. found=421.1.

Example 46 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methoxy-pyrazolo[3,4-d]pyrimidine-1-yl)-ethanone

To a solution of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-d]pyrimidine-1-yl)-ethanone(0.024 mmol, 1 eq.) in 1 mL of MeOH was added solid potassium carbonate(excess), the resultant mixture was heated at 70° C. for 30 minutes,then filtered and dried under vacuum. The crude product was purified byHPLC to provide the desired product as a white powder: ¹H NMR (400 MHz,CDCl₃) δ 8.90 (s, 1H), 8.07 (s, 1H), 7.22 (d, 1H), 6.50 (d, 1H), 6.44(dd, 1H), 5.29 (s, 2H), 3.90 (s, 3H), 3.81 (m, 4H), 3.19 (m, 4H), 2.25(s, 3H). LCMS observed for (M+H)⁺: 418.9.

Example 47 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-2-yl)-ethanone

Preparation of (2,6-dichloro-3-pyridinyl)methanol: To a solution of2,6-dichloro-3-nicotic acid (9 mmol, 1 eq.) in 10 mL of dry THF at 0°C., was added NaBH₄ (27 mmol, 3 eq.) portion by portion under nitrogenatmosphere. After the evolution of hydrogen gas subsided (which isobserved as bubbling in the reaction mixture), BF₃.OMe₂ (27 mmol, 3 eq.)was added dropwise to the reaction mixture at 0° C. The resultantmixture was stirred at 0° C. for 20 minutes followed by the slowaddition of sat. NH₄Cl aq. solution. The reaction solution was thenwarmed to it, and extracted with 300 mL of EtOAc, and the organic layerwas washed with brine, dried over sodium sulfate, filtered andconcentrated in vacuo to provide a white solid, which was used insubsequent reaction without further purification: HPLC retentiontime=0.71 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=178.0. found=178.0.

Preparation of 2,6-dichloro-3-formylpyridine: To a solution of the abovealcohol (2 mmol, 1 eq.) in 10 mL of dichloromethane was added potassiumcarbonate (excess) as a solid, and Dess-Martin periodinate (2 mmol, 1eq.) at rt. The resultant mixture was stirred at it for 30 minutes. A 5%sodium thiosulfate aq. solution was added to the reaction mixture andthe resultant mixture was stirred for another 10 minutes. The reactionmixture was extracted with 300 mL of EtOAc, and the organic layer waswashed with 50 ml of 5% sodium thiosulfate aq. solution, Sat. sodiumbicarbonate aq. solution, brine, and dried over sodium sulfate.Evaporation of solvent in vacuo to provide 200.1 mg of the desiredproduct as a white solid: ¹H NMR (400 MHz, CDCl₃) δ 10.37 (s, 1H), 8.17(d, 1H), 7.42 (d, 1H).

Preparation of 6-chloro-pyrazolo[3,4-b]pyridine: To a solution of2,6-dichloro-3-formylpyridine (0.89 mmol, 1 eq.) in 3 ml THF was addedhydrazine (1.06 mmol, 1.2 eq.) at rt. The resultant solution was heatedat 120° C. in sealed tube for overnight. The solvent was removed invacuo, and the residue was dry loaded on silica gel column. Purificationby silica gel chromatography provide 29.5 mg final product: HPLCretention time=2.17 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35°C.) using 1 ml/min flow rate, a 2.5 minute gradient of 0% to 100% B witha 1.1 minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9%water, B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=154.0. found=154.0.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanoneand1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-2-yl)-ethanone:The two title compounds were synthesized according to the standardcoupling procedure described in Example 6: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-1-yl)-ethanone;¹H NMR (400 MHz, CDCl₃) δ 8.08 (s, 1H), 8.01 (d, 1H), 7.25 (d, 1H), 7.16(d, 1H), 6.50 (d, 1H), 6.45 (dd, 1H), 5.40 (s, 2H), 3.90 (s, 3H), 3.77(m, 4H), 3.23 (m, 4H). LCMS observed for (M+H)⁺: 420.5: For1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-b]pyridine-2-yl)-ethanone:HPLC retention time=1.66 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile). LCMS observed for (M+H)⁺: 420.5.

Example 48 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4-d]pyridine-1-yl)-ethanone

Preparation of 1H-6-hydrazo-pyrazolo[3,4-d]pyridine: To a solution of2,6-dichloro-3-pyridinecarbaldehyde in 2 mL of dioxane was added excessamount of hydrazine. The resultant solution was heated at 150° C.overnight. Upon cooling to rt, the desired product precipitated out ofsolution as a white solid. The crude product was isolated by filtration,washed with a small amount of dioxane, and dried in vacuo. The crudeproduct was used without further purification: HPLC retention time=1.78minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/minflow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute washat 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile); MS (ES) M+H expect=337.9.found=337.9.

Preparation of 1H-6-azido-pyrazolo[3,4-d]pyridine:1H-6-hydrazo-pyrazolo[3,4-d]pyridine was suspended into a mixture of 5mL of concentrated HCl and 10 mL of water at 0° C., and to it was addeddropwise a solution of sodium nitrate in 5 mL of water. The resultantmixture was stirred at 0° C. for 10 min and warmed to rt. The reactionmixture was neutralized to pH=7-8, and extracted with (200 mL×2) EtOAc.The combined organic extract was washed with brine, dried over sodiumsulfate, filtered and concentrated in vacuo to provide the desiredproduct which was used without further purification: HPLC retentiontime=0.50 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=161.0. found=160.8.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4-d]pyridine-1-yl)-ethanone:The title compound was synthesized according to alkylation protocoldescribed in Example 6: HPLC retention time=2.22 minutes (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minutegradient of 20% to 100% B with a 1.1 minute wash at 100% B (A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile); MS (ES) M+H expect=427.1. found=427.1.

Example 49 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-amido-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

To a solution of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4-b]pyridine-1-yl)-ethanone(0.071 mmol, 1 eq.) in 1 mL of EtOAc was added SnCl₂.2H₂O as a solid.The resultant mixture was heated at 40° C. for 2 h. The resultantmixture was cooled to rt and diluted with 200 mL of EtOAc and 50 mL ofSat. sodium bicarbonate aq. solution. The diluted mixture was stirredfor an additional 1 h, before the organic layer was separated, washedwith brine, and dried over sodium sulfate. The solvent was removed invacuo, and the residue was purified by HPLC to provide 10 mg titlecompound: ¹H NMR (400 MHz, CDCl₃) δ 7.86 (s, 1H), 7.74 (d, 1H), 7.24 (d,1H), 7.20 (d, 1H), 6.45 (d, 1H), 6.39 (dd, 1H), 5.16 (s, 2H), 3.88 (s,3H), 3.89 (m, 4H), 3.13 (m, 4H). LCMS observed for (M+H)⁺: 401.1.

Example 50 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-amino-pyrazolo[3,4-c]pyridine-1-yl)-ethanone

Preparation of 7-hydrazo-pyrazolo[3,4-c]pyridine: To a solution of2-chloro-3-fluoro-4-formylpyridine (5.75 mmol, 1 eq.) in 20 mL of THFwas added 1 mL of hydrazine (excess). The resultant solution was heatedat 110° C. in sealed tube for 5 h. The reaction was cooled to rt andsolvent was removed in vacuo. The crude residue was washed several timeswith hexane, EtOAc, and dried in vacuo to provide a light yellow solid,which was used without further purification: HPLC retention time=0.20minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/minflow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute washat 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formicacid/5% water/94.9% acetonitrile); MS (ES) M+H expect=150.1.found=150.0.

Preparation of 7-azido-pyrazolo[3,4-c]pyridine: This compound wassynthesized according to protocol described in Example 48: HPLCretention time=0.26 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35°C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=161.0. found=160.9.

Preparation of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-azido-pyrazolo[3,4-c]pyridine-2-yl)-ethanone:This compound was synthesized according to protocol described in Example6: HPLC retention time=2.43 minutes (Agilent Zorbax SB-C18, 2.1×50 mm,5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to100% B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=427.1. found=427.2.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-amino-pyrazolo[3,4-c]pyridine-1-yl)-ethanone:The title compound was synthesized according to the procedure outlinedin Example 49: ¹H NMR (400 MHz, CDCl₃) δ 7.86 (s, 1H), 7.74 (d, 1H),7.24 (d, 1H), 7.20 (d, 1H), 6.45 (d, 1H), 6.39 (dd, 1H), 5.16 (s, 2H),3.88 (s, 3H), 3.89 (m, 4H), 3.13 (m, 4H). LCMS observed for (M+H)⁺:401.1.

Example 51 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(oxazole-2-yl)-pyrazolo[3,4-b]pyridine-2-yl]-ethanone

Preparation of (3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic acid ethylester: To a solution of 3-Iodo-2H-pyrazolo[3,4-b]pyridine (4 mmol, 1eq.) in 10 mL of dry THF was added dropwise 0.5 M KHMDS (potassiumhexamethyldisilazide) in toluene (4.4 mmol, 1.1 eq.) at −78° C., undernitrogen atmosphere, and the resultant solution was stirred at for 30minutes at −78° C. Chloro ethyl acetate (8 mmol, 2 eq.) was addeddropwise to the reaction solution and the reaction solution was warmedto rt over 1.5 hour and stirred overnight. Following an aqueous workup,the crude product was purified by silica gel chromatography (20% EtOAcin hexane to 70% EtOAc in hexane) to provide 70.2 mg of(3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic acid ethyl ester: HPLCretention time=2.63 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35°C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.08% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=332.0. found=332.1.

Preparation of (3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic acid: Thiscompound was synthesized according to the standard ester hydrolysisprotocol as described in Example 30 using 1N LiOH as the base. The crudeproduct was used in the next step without purification: HPLC retentiontime=1.02 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.08% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=303.0. found=303.5.

Preparation of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridine-2-yl)-ethanone:This compound was synthesized according to standard peptide couplingprotocol using HATU as the coupling reagent: HPLC retention time=297minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/minflow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute washat 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.08%formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=512.0.found=512.5.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(oxazole-2-yl)-pyrazolo[3,4-b]pyridine-2-yl]ethanone:To a mixture of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridine-2-yl)-ethanone(0.071 mmol, 1 eq.) and tetrakis triphenylphosphine palladium (0.025mmol, 0.35 eq.) under nitrogen atmosphere was added 0.5 mL of THF and2-oxazole-(tri-n-butyl) Tin (0.48 mmol, 6.7 eq.). The resultant mixturewas heated in a sealed tube at 80° C. for 48 h. The reaction solutionwas cooled to rt, diluted with 30 mL of NH₄Cl sat. aq. solution, andextracted with 300 mL of EtOAc. The organic layer was separated, washedwith brine, dried over sodium sulfate, filtered and concentrated invacuo. The crude residue was purified by silica gel chromatography (0%to 15% MeOH in EtOAc) to provide 12.3 mg the title compound: ¹H NMR (400MHz, CDCl₃) δ 8.77 (dd, 1H), 8.51 (dd, 1H), 7.81 (d, 1H), 7.22 (m, 3H),6.52 (d, 1H), 6.48 (dd, 1H), 6.01 (s, 2H), 3.90 (s, 3H), 3.80 (m, 4H),3.27 (m, 4H). LCMS observed for (M+H)⁺: 453.5.

Example 52 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4-b]pyridine-1-yl]-ethanone

Preparation of 3-methyl-5-nitro-pyrazolo[3,4-b]pyridine:3-Methyl-pyrazolo[3,4-b]pyridine (1 mmol, 1 eq.) was suspended into amixture of 1:1 fuming nitric acid and concentrated sulfuric acid (1 mL:1mL), and the resultant mixture was heated at 90° C. for 30 minutes. Thereaction mixture was then cooled to rt, and poured into a mixture ofsodium bicarbonate and ice. The resultant solution was warmed up to rtand extracted with 300 mL of EtOAc. The organic extract was separated,washed with brine, dried over sodium sulfate, filtered, and concentratedin vacuo. The crude residue was purified by silica gel chromatography toprovide 70.2 mg of 3-methyl-5-nitro-pyrazolo[3,4-b]pyridine.

Preparation of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-nitro-pyrazolo[3,4-b]pyridine-1-yl]-ethanone:This compound was synthesized from3-methyl-5-nitro-pyrazolo[3,4-b]pyridine according to the alkylationprotocol described in Example 6: HPLC retention time=1.46 minutes(Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate,a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100% B(A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%water/94.9% acetonitrile); MS (ES) M+H expect=445.1. found=445.1.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4-b]pyridine-1-yl]-ethanone:1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-nitro-pyrazolo[3,4-b]pyridine-1-yl]ethanone(15 mg) is combined with 200 mg of iron powder in 2 mL of acetic acid at100° C. for 30 min. After cooling to rt, the reaction solution wasdiluted with EtOAc and filtered. The filtrate was evaporated in vacuoand purified by HPLC to provide1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4-b]pyridine-1-yl]-ethanone:HPLC retention time=1.46 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ,35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% Bwith a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile); MS (ES) M+H expect=414.2. found=415.1.

Example 53 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-amino-6-methyl-pyrazolo[3,4-b]pyridine-1-yl]-ethanone

Preparation of 1H-3-amino-6-methyl-pyrazolo[3,4-d]pyridine: Thiscompound was synthesized according to the cyclization procedure usinghydrazine described in Example 3 and the crude product was used in thenext step without further purification.

Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-amino-6-methyl-pyrazolo[3,4-b]pyridine-1-yl]-ethanone:This compound was synthesized according to the standard couplingprocedure described in Example 6: ¹H NMR (400 MHz, CDCl₃) 7.75 (d, 1H),7.22 (d, 1H), 6.86 (d, 1H), 6.48 (d, 1H), 6.42 (dd, 1H), 5.18 (s, 2H),3.89 (s, 3H), 3.75 (m, 4H), 3.16 (m, 4H), 2.62 (s, 3H). LCMS observedfor (M+H)⁺: 415.5.

Example 54 Synthesis of1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)ethanone

Preparation of1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:A mixture of2-Chloro-1-[(S)-4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-ethanone(1.37 g, 4.08 mmol, 1 eq), 3-Iodo-1H-pyrazolo[3,4-b]pyridine (1.0 g,4.08 mmol, 1 eq), potassium carbonate (2.26 g, 16.4 mmol, 4 eq), and DMF(15 ml) was stirred overnight at 90° C. The reaction solution wasdiluted with ethyl acetate, washed with saturated aqueous NaHCO₃, andconcentrated in vacuo. The crude product was purified by flashchromatography to provide1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(2.2 g).

Preparation of1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile:A mixture of1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(2.2 g, 4.0 mmol, 1 eq), CuCN (3.6 g, 40 mmol, 10 eq), and DMF (25 ml)was stirred at 175° C. for 1 hrs. The reaction mixture was cooled to rt,diluted with ethyl acetate and filtered. The filtrate was washed withwater, dried over Na₂SO₄, and purified by flash chromatography toprovide1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile(1.6 g).

Preparation of1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine:A mixture of1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile(1.6 g, 3.6 mmol, 1 eq), NH₂OH.HCl (0.84 g, 10.8 mmol, 3 eq), TEA (1.5ml), and ethanol (10 ml) was stirred at 65° C. overnight. The reactionsolution was concentrated in vacuo, and dissolved in ethyl acetate,washed with brine, and concentrated to provide1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine(1.2 g).

Preparation of1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:A mixture of1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine(1.2 g), trimethyl orthoformate (20 ml) and para-toluene sulfonic acid(PTSA) (0.1 g) was stirred at 100° C. overnight. The reaction mixturewas concentrated in vacuo to provide a crude residue which was purifiedby flash chromatography to provide1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(0.7 g). LCMS Retention time: 2.61 min (Agilent Zorbax SB-C18, 2.1×50mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20%to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%acetonitrile). LCMS observed for (M+H)⁺: 486.

Example 55 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

A solution of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4-b]pyridine-1-yl)-ethanone(0.15 mmol, 1 eq.) and hydroxylamine HCl salt (0.45 mmol, 3 eq.) in 2.5mL of EtOH was heated at 60° C. for 1 h. The reaction mixture was cooledto rt, and concentrated in vacuo. The crude residue was dissolved with200 mL of dichloromethane, washed with 50 mL of 5% K₂CO₃ aq. solution,brine solution, dried over sodium sulfate, filtered and concentrated invacuo to provide the desired product as a white solid: HPLC retentiontime=1.61 minutes (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C.) using 1ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water,B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+Hexpect=444.1. found=444.5.

Example 56 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(oxadiazole-3-yl)-pyrazolo[3,4-b]pyridine-1-yl]-ethanone

To a suspension of (0.067 mmol, 1 eq.) of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4-b]pyridine-1-yl)-ethanonein 2 mL of trimethylorthoformate was added camphorsulfonic acid (CSA)(5.0 mg, catalytic amount). The resultant mixture was heated at 50° C.for 10 minutes and cooled to rt. The reaction solution was concentrationin vacuo to provide a crude residue which was purified by HPLCchromatography to provide 20.0 mg of the title compound: ¹H NMR (400MHz, CDCl₃) 8.83 (s, 1H), 8.62 (dd, 1H), 7.35 (dd, 1H), 7.22 (d, 1H),6.51 (d, 1H), 6.44 (dd, 1H), 5.59 (s, 2H), 3.89 (s, 3H), 3.79 (m, 4H),3.23 (m, 4H). LCMS observed for (M+H)⁺: 454.5.

Example 57 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(5-methyl-oxadiazole-3-yl)-pyrazolo[3,4-b]pyridine-1-yl]-ethanone

The title compound was synthesized according to the cyclizationprocedure using trimethylorthoacetate as described in Example 56: ¹H NMR(400 MHz, CDCl₃) δ 8.61 (dd, 1H), 7.31 (dd, 1H), 7.22 (d, 1H), 6.54 (d,1H), 6.42 (dd, 1H), 5.57 (s, 2H), 3.89 (s, 3H), 3.77 (m, 4H), 3.21 (m,4H), 2.69 (s, 1H). LCMS observed for (M+H)⁺: 468.5.

Example 58 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-acetimido-pyrazolo[3,4-b]pyridine-1-yl)-ethanone

1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-(6-amido-pyrazolo[3,4-b]pyridine-1-yl)-ethanone,acetic anhydride (1.2 equiv) and pyridine (3 equiv) was combined in DCMat rt for 30 min: HPLC retention time=1.82 minutes (Agilent ZorbaxSB-C18, 2.1×50 mm, 5μ, 35° C.) using 1 ml/min flow rate, a 2.5 minutegradient of 20% to 100% B with a 1.1 minute wash at 100% B (A=0.1%formic acid/5% acetonitrile/94.9% water, B=0.08% formic acid/5%water/94.9% acetonitrile); MS (ES) M+H expect=443.1. found=442.8.

Example 59 Synthesis of1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methylsulfonyl-pyrazolo[4,3-c]pyridine-1-yl)-ethanone

The title compound was synthesized from1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanoneaccording to the protocol described in Example 41: ¹H NMR (400 MHz,CDCl₃) 8.65 (d, 1H), 8.48 (d, 1H), 7.39 (dd, 1H), 7.22 (d, 1H), 6.51 (s,1H), 6.44 (d, 1H), 5.53 (s, 2H), 3.91 (s, 3H), 3.78 (m, 4H), 3.34 (s,3H), 3.22 (m, 4H), LCMS observed for (M+H)⁺: 415.0.

Example 60 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone

Preparation of Ethyl (3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate: To amixture of 3-iodo-1H-pyrazolo[3,4-b]pyridine (9.8 g, 40 mmol, 1 equiv)and potassium carbonate (27.6 g, 5 equiv) in 15 mL of DMF at 90° C. wasadded ethyl chloroacetate (8.5 mL, 40 mmol, 1 equiv). Two hours later,the reaction mixture was diluted with ethyl acetate followed by washingwith saturated aqueous NaHCO₃. The organic layer was dried andconcentrated to provide the crude product. Purification of the crudeproduct by flash chromatography gave ethyl(3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate (11 g).

Preparation of (3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid: Ethyl(3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate (11 g, 33 mmol, 1 equiv)was dissolved in 50 mL of THF and 50 mL of MeOH to the solution wasadded 40 mL of 1N LiOH for 3 h. The organic solvents were evaporated andthe remaining aqueous phase was neutralized with 1N HCl to a pH of about1 which resulted in the precipitation of the desired product as a whitesolid was filtered and air dried to give(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid.

Preparation of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:A mixture of (3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid (3.03 g,10 mmol, 1 equiv), 1-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazine(2.45 g, 1 equiv), BOP reagent (4.86 g, 1 equiv), triethylamine (4.2 mL,3 equiv) in 10 mL of MF was stirred at rt overnight. To the reactionmixture was then added water and the solid precipitates were removed byfiltration and air dried to give1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone.LCMS (ES) observed for M+H 530.0.

Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone:To a solution of oxazole (690 mg 10 mmol, 2.5 equiv) in tetrahydrofuran(5 mL) under nitrogen atmosphere, was added dropwise n-butyl lithium(2.5 M in Hexane, 4.8 mL, 3 equiv.). The resultant mixture was stirredat −78° C. for an additional 60 min followed by the addition of ZnCl₂(0.5 M in THF, 32 mL, 4 equiv.). The reaction solution was allowed towarm to 0° C. and stirred 1 h followed by the addition of1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(2.12 g, 4 mmol, 1 equiv) and palladium tetrakis(triphenylphosphine)(462 mg, 0.1 equiv). The reaction mixture was then heated to reflux for12 hr, cooled to room temperature and diluted with ethyl acetate. Thereaction mixture was washed with water, brine, dried over sodiumsulfate, and concentrated in vacuo to provide the crude product.Purification by flash chromatography provided of the desired product1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanoneas a white powder (1.03 g). LCMS (ES) observed for M+H 471.1. HPLCretention time=2.4 min (Agilent Zorbax SB-C18, 2.1×50 mm, 5μ, 35° C., 1mL/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 minwash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%formic acid/5% water/94.9% acetonitrile).

Example 61 Synthesis of1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

Step 1: A mixture of 3-iodo-7-azaindazole (25.50 g) and K₂CO₃ (41.4 g)in DMF (200 mL) was heated to 85° C. and t-butyl chloroacetate (14.3 mL)was slowly added. The mixture was stirred at this temperature for 1 hour(h), cooled to room temperature followed by the addition of water (300mL). Filtration of the reaction mixture provided(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester.

Step 2: A 250 mL flask was charged with(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester (15.0g), PdCl₂(dppf) (3.0 g), Zn(CN)₂ (4.96 g), DMF (200 mL) and H₂O (14 mL).The flask containing the resultant suspension was degassed andbackfilled with nitrogen gas repeatedly for 5 minutes, followed byaddition of Pd₂(dba)₃ (3.85 g) to the reaction mixture. The reactionmixture was heated under N₂ at 90° C. for 16 h, cooled to roomtemperature, diluted with H₂O (800 mL) and filtered. The collected solidwas washed with toluene (10 mL) to provide(3-cyano-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester as ayellow solid.

Step 3: A mixture of (3-cyano-pyrazolo[3,4-b]pyridin-1-yl)-acetic acidtert-butyl ester, hydroxylamine hydrochloride (8.28 g) and Et₃N (22.6mL) in EtOH (120 mL) was heated over night under N₂ at 65° C. Themixture was cooled to room temperature, filtered and the collected solidwas washed with H₂O (100 mL) and Et₂O (50 mL×2) to afforded[3-(N-hydroxycarbamimidoyl)-pyrazolo[3,4-b]pyridin-1-yl]-acetic acidtert-butyl ester.

Step 4: [3-(N-Hydroxycarbamimidoyl)-pyrazolo[3,4-b]pyridin-1-yl]-aceticacid tert-butyl ester (6.17 g) in a 100 mL vial was charged with AcOH(45 mL) and Ac₂O (4.3 mL). The resultant mixture was stirred at roomtemperature for 1 h at this time the initial suspension became a clearsolution. To this solution was added Pd/C (10%, 900 mg) and stirredunder 1 atm H₂ balloon, and resultant mixture was stirred over night atroom temperature. Filtration from celite washed with DCM/MeOH andevaporation gave (3-amidino-pyrazolo[3,4-b]pyridin-1-yl)-acetic acidtert-butyl ester which was used without further purification.

Step 5: (3-Amidino-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butylester obtained above in a 100 mL vial was charged withchloroacetylaldehyde (5.72 mL), dioxane (50 mL) and K₂CO₃ (12.42 g). Theresultant mixture was stirred at 80° C. for 4 h and morechloroacetylaldehyde (5.72 mL) and K₂CO₃ (12.42 g) were added. Themixture was stirred another 1 h at 80° C. and stirred at 120° C. foranother 1 h, cooled to room temperature, diluted with dichloromethane(DCM), washed with brine, dried (Na₂SO₄), filtered and evaporated invacuo. Purification by flash chromatography provided2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acidtert-butyl ester as a brown oil.

Step 6: 2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)aceticacid tert-butyl ester (977 mg) was dissolved in trifluoroacetic acid(TFA) (10 mL) and stirred at room temperature for 1 hr. The mixture wasevaporated in vacuo to provide2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid as abrown oil, which was used without further purification.

Step 7: (Protocol A—the HBTU coupling procedure) A solution of[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]acetic acid (0.30 M,0.40 mL, 0.12 mmol) was transferred to a vial.1-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazine dihydrochloride (48 mg,0.14 mmol), 0-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HBTU) (55 mg, 0.14 mmol) and i-Pr₂NEt (0.30 mL)were added to the vial and the mixture was stirred at ambienttemperature. After 30 minutes, LC/MS analysis indicated formation ofdesired product and complete consumption of the carboxylic acid startingmaterial. The mixture was diluted with EtOAc, washed with water (1×) andbrine (1×), dried over Na₂SO₄ and evaporated. The residue was purifiedby silica gel chromatography (1% to 8% MeOH in CH₂Cl₂) to provide1-[4-(4-chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.79 (dd, 0.6; H, J=8.4, 1.6Hz), 8.66 (dd, 0.4; H, J=8.0, 1.6 Hz), 8.57-8.55 (m, 1H), 7.29-7.26 (m,1H), 7.22-7.16 (m, 1H), 7.09-7.05 (m, 2H), 6.50-6.45 (m, 2H), 5.45 (s,0.6H), 5.43 (s, 1.4H), 4.07-4.01 (m, 2H), 3.81-3.69 (m, 4H), 3.17-3.13(m, 1.6H), 3.08-3.02 (m, 2.4H), 1.50-1.42 (m, 3H); LC/MS m/z (M+H)⁺484.4.

Example 62 Synthesis of1-{4-[4-Chloro-2-fluoro-5-(2-fluoroethoxy)-phenyl]piperazin-1-yl}-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.1-[4-Chloro-2-fluoro-5-(2-fluoroethoxy)phenyl]-piperazinedihydrochloride and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (2% to 3.5% MeOH in CH₂Cl₂) to provide1-{4-[4-chloro-2-fluoro-5-(2-fluoroethoxy)phenyl]piperazin-1-yl}-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.79 (dd, 0.6; H, J=8.0, 1.8Hz), 8.67 (d, 0.4; H, J=6.4 Hz), 8.57-8.55 (m, 1H), 7.30-7.25 (m, 1H),7.11-7.06 (m, 2H), 6.63 (d, 0.6H, J=7.6 Hz), 6.57 (d, 0.4H, J=7.6 Hz),6.54 (d, 1H, J=7.6 Hz), 5.45 (s, 0.7H), 5.43 (s, 1.3H), 4.83-4.80 (m,1H), 4.71-4.68 (m, 1H), 4.31-4.18 (m, 2H), 3.82-3.76 (m, 3H), 3.50 (t,1H, J=5.2 Hz), 3.23-3.04 (m, 4H); LC/MS m/z (M+H)⁺ 502.4.

Example 63 Synthesis of1-[4-(4-Chloro-3-ethoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.1-(4-Chloro-3-ethoxyphenyl)piperazine dihydrochloride and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (4% to 15% MeOH in CH₂Cl₂) to provide1-[4-(4-chloro-3-ethoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid (25 mg): ¹H NMR (CDCl₃, 400 MHz) δ 8.79 (dd, 0.6H), 8.66(dd, 0.4H), 8.57-8.54 (m, 1H), 7.29-7.19 (m, 4H), 6.49-6.40 (m, 2H),5.45 (s, 0.7H), 5.43 (s, 1.3H), 4.10-4.04 (m, 2H), 3.81-3.69 (m, 4H),3.23-3.16 (m, 4H), 1.50-1.45 (m, 3H); LC/MS m/z (M+H)⁺ 466.4.

Example 64 Synthesis of1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.1-(4-Chloro-2-fluoro-5-methoxyphenyl)piperazine dihydrochloride and[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (1% to 10% MeOH in CH₂Cl₂) to provide1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid (27 mg): ¹H NMR (CDCl₃, 400 MHz) δ 8.79 (dd, 0.6H), 8.67(dd, 0.4H), 8.57-8.55 (m, 1H), 7.31-7.20 (m, 2H), 7.12-7.06 (m, 2H),6.49-6.45 (m, 1H), 5.45 (s, 0.6H), 5.43 (s, 1.4H), 3.86 (s, 0.9H), 3.85(s, 2.1; H), 3.81-3.75 (m, 4H), 3.15-3.08 (m, 4H); LC/MS m/z (M+H)⁺470.4.

Example 65 Synthesis of1-[(S)-4-(4-Chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.(S)-1-(4-Chloro-3-methoxyphenyl)-3-methylpiperazine dihydrochloride and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (1% to 8% MeOH in CH₂Cl₂) to provide1-[(S)-4-(4-chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.79 (dd, 0.6H), 8.66 (dd,0.4H), 8.57-8.55 (m, 1H), 7.29-7.19 (m, 4H), 6.44-6.39 (m, 2H), 5.42(br. s, 2H), 4.83 (br. s, 0.3H), 4.49 (br. s, 0.3H), 4.30 (br. s, 0.3H),3.89 (s, 1.2H), 3.88 (s, 1.8H), 3.83 (br. s, 0.3H), 3.72-3.69 (m, 1H),3.54-3.52 (m, 1H), 3.38 (br. s, 1H), 3.19-3.15 (m, 1H), 3.00 (br. s,1H), 2.80 (br. s, 1H), 1.50-1.43 (m, 3H); LC/MS m/z (M+H)⁺ 466.4.

Example 66 Synthesis of1-[(S)-4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following protocol A.(S)-1-(4-Chloro-2-fluoro-5-methoxy-phenyl)-3-methylpiperazinedihydrochloride and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (2% to 3% MeOH in CH₂Cl₂) to provide1-[(S)-4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid (29 mg): ¹H NMR (CDCl₃, 400 MHz) 8.79 (dd, 1H), 8.55 (dd,1H), 7.27-7.21 (m, 3H), 7.07 (d, 1H), 6.43 (br. d, 1H), 5.46-5.37 (m,2H), 4.83 (br. s, 0.3H), 4.51-4.48 (m, 0.6H), 4.28-4.21 (m, 0.6H), 3.86(s, 0.9H), 3.85 (s, 2.1H), 3.79 (br. s, 0.3H), 3.67 (br. s, 0.3H),3.33-3.21 (m, 2.5H), 2.95-2.93 (m, 0.9H), 2.83-2.76 (m, 1.6H), 1.48-1.40(m, 3H); LC/MS m/z (M+H)⁺ 484.4.

Example 67 Synthesis of1-[(R)-4-(4-Chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.(R)-1-(4-Chloro-3-methoxyphenyl)-2-methylpiperazine and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (1% to 7.5% MeOH in CH₂Cl₂) to provide1-[(R)-4-(4-chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.76 (d, 0.6H), 8.66 (dd,0.3H), 8.57-8.54 (m, 1H), 7.29-7.19 (m, 4H), 6.48-6.40 (m, 2H),5.53-5.40 (m, 2H), 4.26 (br. d, 0.6H), 4.00 (br. d, 0.6H), 3.88 (s,1.3H), 3.86 (s, 1.7H), 3.80-3.49 (m, 3.2H), 3.33 (br. s, 0.6H),3.17-3.14 (m, 2H), 1.51-1.42 (m, 3H); LC/MS m/z (M+H)⁺ 466.4.

Example 68 Synthesis of1-[(S)-4-(4-Chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following protocol A.(S)-1-(4-Chloro-3-methoxyphenyl)-2-methylpiperazine and[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (1% to 7% MeOH in CH₂Cl₂) to provide1-[(S)-4-(4-chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.77 (d, 0.6H), 8.66 (d,0.3H), 8.57-8.54 (m, 1H), 7.30-7.19 (m, 4H), 6.48-6.40 (m, 2H),5.54-5.36 (m, 2H), 4.25 (br. d, 0.6H), 4.00 (br. d, 0.6H), 3.88 (s,1.3H), 3.86 (s, 1.7H), 3.82-3.48 (m, 3.2H), 3.36-3.29 (m, 0.6H),3.17-3.13 (m, 2H), 1.51-1.43 (m, 3H); LC/MS m/z (M+H)⁺ 466.4.

Example 69 Synthesis of1-[(R)-4-(4-Chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

The title compound was prepared following Protocol A.(R)-1-(4-Chloro-3-methoxyphenyl)-3-methylpiperazine and[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]acetic acid were usedas the coupling components. The crude product was purified by silica gelchromatography (1% to 7.5% MeOH in CH₂Cl₂) to provide1-[(R)-4-(4-chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a tan solid: ¹H NMR (CDCl₃, 400 MHz) δ 8.75 (d, 0.6H), 8.66 (dd,0.4H), 8.57-8.54 (m, 1H), 7.29-7.18 (m, 4H), 6.44-6.39 (m, 2H), 5.42(br. s, 2H), 4.82 (br. s, 0.3H), 4.45 (br. s, 0.3H), 4.33 (br. s, 0.3H),3.88 (s, 1.2H), 3.87 (s, 1.8H), 3.83 (br. s, 0.3H), 3.73-3.67 (m, 1H),3.54-3.52 (m, 1H), 3.38 (br. s, 1H), 3.17-3.13 (m, 1H), 2.99 (br. s,1H), 2.80 (br. s, 1H), 1.50-1.42 (m, 3H); LC/MS m/z (M+H)⁺ 466.4.

Example 70 Synthesis of2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-((S)-4-(4-chloro-3-ethoxyphenyl)-2-methylpiperazin-1-yl)ethanone

The title compound was prepared following Protocol A. To a vialcontaining (S)-1-(4-chloro-3-ethoxyphenyl)-3-methylpiperazinedihydrochloride (70 mg, 0.21 mmol) was added2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (51mg, 0.21 mmol), HBTU (81 mg, 0.21 mmol), DMF (0.7 mL), and DIPEA (0.15mL, 0.87 mmol). The reaction mixture was maintained at 30° C. for 24 h.The solution was diluted with EtOAc (30 mL) and washed with 1N HCl (2×10mL) and sat. aq. NaCl (2×10 mL). The organic phase was dried over MgSO₄and concentrated in vacuo. The resulting residue was purified bypreparative HPLC (20→95% gradient of MeCN—H₂O with 0.1% TFA) and thepure fractions lyophilized to afford the indicated compound (11 mg, 11%yield): MS (ES) [M+H]⁺ expected 480.2. found 480.5; ¹H NMR (CDCl₃, 400MHz) δ 9.94 (br s, 1H), 8.79 (dd, J=1.6, 8.0, 1H), 8.55 (dd, J=1.6, 4.4,1H), 7.19-7.26 (m, 4H), 6.37-6.43 (m, 2H), 5.40 (br s, 2H), 2.78-4.81(m, 10H), 4.07 (q, J=6.8, 2H), 1.46 (t, J=6.8, 3H).

Example 71 Synthesis of2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(3-(2-fluoroethoxy)-4-chlorophenyl)piperazin-1-yl)ethanone

This compound was prepared by Protocol A. To a vial containing1-(3-(2-fluoroethoxy)-4-chlorophenyl)piperazine dihydrochloride (70 mg,0.21 mmol) was added2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (51mg, 0.21 mmol), HBTU (83 mg, 0.22 mmol), DMF (0.7 mL), and DIPEA (0.20mL, 1.2 mmol). The reaction mixture was maintained at 20° C. for 24 h.The solution was diluted with EtOAc (30 mL) and washed with 1N HCl (2×10mL) and sat. aq. NaCl (2×10 mL). The organic phase was dried over MgSO₄and concentrated in vacuo. The resultant residue was purified bypreparative HPLC (20→95% gradient of MeCN—H₂O with 0.1% TFA) and thepure fractions lyophilized to afford the indicated compound (20 mg, 20%yield): MS (ES) [M+H]⁺ expected 484.2. found 484.4. ¹H NMR (CDCl₃, 400MHz) δ 9.94 (br s, 1H), 8.78 (dd, J=1.2, 8, 1H), 8.55 (dd, J=1.2, 4.6,1H), 7.16-7.26 (m, 4H), 6.45-6.53 (m, 2H), 5.44 (s, 2H), 4.77 (dt,J=4.0, 46.8, 2H), 4.26 (dt, J=4.0, 26.8, 2H), 3.69-3.78 (m, 4H),3.10-3.21 (m, 4H).

Example 72 Synthesis of2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-((S)-4-(3-(2-fluoroethoxy)-4-chlorophenyl)-2-methylpiperazin-1-yl)ethanone

This compound was prepared by Protocol A. To a vial containing(S)-1-(3-(2-fluoroethoxy)-4-chlorophenyl)-3-methylpiperazinedihydrochloride (80 mg, 0.23 mmol) was added2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (51mg, 0.21 mmol), HBTU (81 mg, 0.21 mmol), DMF (0.7 mL), and DIPEA (0.2mL, 1.2 mmol). The reaction mixture was maintained at 30° C. for 24 h.The solution was diluted with EtOAc (30 mL) and washed with 1N HCl (2×10mL) and sat. aq. NaCl (2×10 mL). The organic phase was dried over MgSO₄and concentrated in vacuo. The resultant residue was purified bypreparative HPLC (20→95% gradient of MeCN—H₂O with 0.1% TFA) and thepure fractions lyophilized to afford the indicated compound (14 mg, 13%yield): MS (ES) [M+H]⁺ expected 498.2. found 498.4. ¹H NMR (CDCl₃, 400MHz) δ 9.95 (br s, 1H), 8.79 (dd, J=1.6, 8.2, 1H), 8.55 (dd, J=1.6, 4.4,1H), 7.19-7.26 (m, 4H), 6.44-6.50 (m, 2H), 5.40 (br s, 2H), 4.77 (dt,J=4.2, 47.2, 2H), 4.26 (dt, J=4.2, 27.2, 2H), 2.78-4.42 (m, 10H).

Example 73 Synthesis of1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone

This compound was prepared by Protocol A. A vial was charged with2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (55mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),1-(4-Chloro-3-methyl-phenyl)-piperazine dihydrochloride (142 mg, 0.50mmol), anhydrous DMF (2.0 mL), and DIPEA (0.5 mL). The vial was capped,heated to 45° C., and stirred overnight. The following day, thevolatiles were removed in vacuo and separation by preparative hplc(reverse phase, acetonitrile-water gradient) gave1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone:MS (ES) [M+H]⁺ found: 436.4

Example 74 Synthesis of1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone

This compound was prepared by Protocol A. A vial was charged with2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (55mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),1-(4-Chloro-3-trifluoromethyl-phenyl)-piperazine dihydrochloride (170mg, 0.50 mmol), anhydrous DMF (2.0 mL), and DIPEA (0.5 mL). The vial wascapped, heated to 45° C., and stirred overnight. The following day, thevolatiles were removed in vacuo and separation by preparative hplc(reverse phase, acetonitrile-water gradient) gave1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone:MS (ES) [M+H]⁺ found: 490.4

Example 75 Synthesis of1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone

This compound was prepared by Protocol A. A vial was charged with2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid (55mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),1-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazine dihydrochloride (177mg, 0.50 mmol), anhydrous DMF (2.0 mL), and DIPEA (0.4 mL). The vial wascapped, heated to 45° C., and stirred overnight. The following day, thevolatiles were removed in vacuo and separation by preparative hplc(reverse phase, acetonitrile-water gradient) gave1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone.MS (ES) [M+H]⁺ found: 506.4

Example 76 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone

The title compound was prepared from1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-iodo-pyrazolo[3,4-b]pyridin-1-yl]-ethanoneaccording to the procedure similar to those described from step 2 tostep 5 in the synthesis of Example 61: ¹H NMR (CDCl₃, 400 MHz) δ 10.22(br, 1H), 8.82 (dd, 1H), 8.56 (dd, 1H), 7.20-7.30 (m, 3H), 7.11 (s, 1H),6.47 (d, 1H), 6.42 (dd, 1H), 5.44 (s, 2H), 3.88 (s, 3H), 3.80 (m, 4H),3.19 (m, 4H); MS (ES) M+H expect 452.2.

Example 77 Synthesis of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone

To the solution of1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone(50 mg, 0.11 mmol, 1 eq) in THF was added 60% sodium hydride (5.7 mg,0.14 mmol, 1.3 eq) and stirred for 1 hr, followed by the addition ofiodomethane (25.8 mg, 0.16 mmol, 1.5 eq). 2 hrs later, LCMS indicatedthat the major peak is the desired product preparative hplc (reversephase, acetonitrile-water gradient) gave1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone.MS (ES) [M+H]⁺ found: 465.2.

Example 78

Step 1: Tert-butyl 4-(4-chloro-3-methoxyphenyl)piperazine-1-carboxylate

To a 3-necked, 5-L Morton flask equipped with a mechanical stirrer, gasadapter, heating mantle and thermometer was added rac-BINAP (4.24 g,0.005 equiv) and Pd₂(dba)₃ (3.20 g, 0.0025 equiv). The flask wasevacuated, and back-filled with nitrogen. Toluene (100 mL) was added bycannula. The mixture was stirred at room temperature for 15 min to givea purple solution. Toluene (2.0 L) was then added.2-Chloro-5-bromoanisole (300.3 g, 1.356 mol, 1 equiv) was added in oneportion. Boc-piperazine (252.4 g, 1 equiv) was added in one portion.Sodium tert-butoxide (183.0 g, 1.4 equiv) was added in one portion. Theflask was evacuated and back-filled with nitrogen. The mixture was thenheated to an internal temperature of 60° C. A heterogeneous light-orangeslurry was obtained. After 1 h, the mixture becomes a homogeneous brownsolution. After an additional 15 h, the mixture was cooled to roomtemperature. EtOAc (2.0 L) was added to the stirring mixture. The solidwas filtered. The filtrate was washed with EtOAc (100 mL). The combinedfiltrate was washed with 10% aq. K₂CO₃ solution (1×1 L), water (1×1 L),and dried over MgSO₄. The solvent was removed in vacuo to afford theproduct as an orange solid (410.3 g, 93% yield).

Step 2: 1-(4-chloro-3-methoxyphenyl)piperazine dihydrochloride

A 4-L beaker equipped with a mechanical stirrer was charged withtert-butyl 4-(4-chloro-3-methoxyphenyl)piperazine-1-carboxylate (500 g,1.53 mol, 1 equiv) and MeOH (1.50 L). While stirring at room temperatureconc. 37% HCl (500 mL, 4 equiv) was added over 5 min. The internaltemperature rose to 40° C., and the solution became thick withprecipitate. After 15 min, the mixture was heated to an internaltemperature of 60° C. on a hotplate. (Foaming begun at approximately 50°C. as the mixture warms.) After 2 h at 60° C., the solution was cooledto room temperature, and subsequently to 5° C. in a refrigerator. Theproduct was collected by filtration in two batches. Each batch of thered-brown filtrate was washed with EtOAc (2×500 mL) to give a lightyellow solid. The two batches were combined to afford the product (391.3g). The filtrate was concentrated to a volume of 300 mL in vacuo andtreated with hot (50° C.) MeOH (500 mL). The mixture was cooled to 5° C.in a refrigerator for 24 h. The resulting precipitate was collected byfiltration and washed with EtOAc (2×200 mL) to afford an additional 44.3g of product (total of 435.6 g, 95% yield).

Step 3: 2-chloro-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanone

To a 3 L flask equipped with a mechanical stirrer was added1-(4-chloro-3-methoxyphenyl)piperazine dihydrochloride (220 g, 0.73 mol,1 equiv), CH₂Cl₂ (1000 mL), and water (1000 mL). The biphasic mixturewas cooled to 5° C. with an ice-water bath. K₂CO₃ (506 g, 5 equiv) wasadded to the vigorously stirring solution in portions to minimizefoaming. A solution of chloroacetyl chloride (124.4 g, 1.5 equiv) inCH₂Cl₂ (100 mL) was added dropwise from an addition funnel, whilemaintaining an internal temperature below 8° C. After 1 h, the coolingbath was removed, and the reaction warmed to room temperature. After anadditional 1 h, the layers were partitioned. The aqueous phase wasextracted with CH₂Cl₂ (2×300 mL), and the combined organic layers driedover 3:1 Na₂SO₄/K₂CO₃ (addition of K₂CO₃ helps the solution phase tobecome clear). After filtration, the filtrate was concentrated in vacuo,and the residue was dried for 16 h under vacuum to afford the product asan off-white solid (410 g, 92% yield).

Step 6: 7-azaindazole-3-carboxaldehyde

A 5 L 3-necked flask equipped with a digital thermometer, a 1 L additionfunnel and mechanical stirrer (all glassware dried in oven and cooled inair for 30 min before use) was charged with 3-iodo-7-azaindazole (196.0g, 0.80 mol) and 1 L of anhydrous THF (in SureSeal bottle from Aldrichand used as is). The solids completely dissolved in THF at roomtemperature to form a dark brown solution. The flask was then cooled to−5° C. with an ice/NaCl bath and moderate stirring and o-tolylmagnesiumchloride (1 M solution in THF, 880 mL, 1.1 equiv) was added dropwise tokeep the internal temperature between −5° C. to −3° C. (after ˜820 mL ofo-tolylmagnesium chloride solution was added, the temperature no longerrose). The whole addition process took 2 hr and 25 min. At the end ofthe addition, the mixture was a homogeneous dark brown solution.

After an additional 1 hr, isopropylmagnesium chloride solution (2 M inTHF, 480 mL, 1.2 equiv) was added dropwise to keep internal temperature<4° C. After 25 min and about 200 mL isopropylmagnesium chloridesolution was added, brown precipitate started to form. After a total of380 mL of isopropylmagnesium chloride solution was added, the mixturebecame homogeneous again. The whole addition process was done in 45 min.After another 1 hr 25 min, a small amount of sample was taken out andquenched with D₂O. LCMS analysis of this sample indicated the completeIodo-Mg exchange.

1-Formylpiperidine (120 mL, 1.3 equiv) was then added dropwise to keepthe internal temperature between <2° C. After about 30 mL1-formylpiperidine was added, the internal temperature did not go upanymore and rest of the 1-formylpiperidine was added relatively quickly.The whole addition process took 20 min. At the end of the addition, themixture was still a dark homogeneous solution and was allowed to slowlywarm up to room temperature and moderately stirred for 18 hr.

The mixture was re-cooled to 0° C. with an ice/NaCl bath and quenched byslow addition of a mixture of saturated NH₄Cl solution (750mL)/concentrated HCl solution (250 mL) to keep the internal temperatureat <35° C. After the addition was complete, stirring was allowed tocontinue for 1 hr and a yellow precipitate appeared. The mixture wasfiltered and the solid was washed with TIM (100 mL). The collectedfiltrate was transferred to a separation funnel and the pH of theaqueous layer was adjusted to between 5 and 6 with the addition ofNaHCO₃ (around 5 g). The THF layer was separated and washed with sat.NaCl solution (2×100 mL). The combined aqueous layers (including theNaCl wash and quenched aqueous layer) was extracted with EtOAc (3×250mL). The combined organic layers was dried (Na₂SO₄), filtered andevaporated in vacuo (bath temperature <30° C.) to give a brownish solid.This solid was triturated with Et₂O (600 mL) and filtered. The collectedsolid was washed with Et₂O (2×100 mL) to give7-azaindazole-3-carboxaldehyde as a yellowish solid (86.6 g, 73%).

Step 4:1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl-pyrazolo[3,4-b]pyridin-1-yl]ethanone

A mixture of 7-azaindazole-3-carboxaldehyde (86.6 g, 0.59 mol, 1 equiv),NaI (8.8 g, 0.1 equiv) and K₂CO₃ (162.5 g, 2 equiv) in DMF (0.5 L) in a5 L flask was heated to 85° C. (the heating process took around 1.5 h).2-chloro-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanone (175 g,1 equiv) was added in small portions to the reaction mixture. The wholeaddition process took about 30 min. The mixture was then stirred at 85°C. for 30 min and LCMS confirmed that the reaction was complete. Aftercooling down to room temperature, the mixture was transferred to a 4 Lflask with 2 L ice. The reaction flask was rinsed with small amount ofacetone (30 mL) and transferred to the DMF/ice mixture in the 4 L flaskalso. A lot of brownish solids precipitated out. After the icecompletely melted, the mixture was filtered. The collected solid waswashed with water (1 L), blended, and then washed with water (1 L) toget rid of some residual DMF. The collected solid contained a lot ofwater, so it was dissolved in CH₂Cl₂ (4 L) and the mixture wastransferred to a 5 L separation funnel. The bottom CH₂Cl₂ layer wasseparated and the top aq. layer was washed with CH₂Cl₂ (2×100 mL). Thecombined CH₂Cl₂ layers were dried (Na₂SO₄), filtered and evaporated invacuo to give1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas a brownish solid (236.4 g, 97%) which was used without purification.

Step 5:1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone

1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl-pyrazolo[3,4-b]pyridin-1-yl]ethanone(300 g, 723 mmol, 1 equiv), glyoxal trimer dihydrate (60.6 g, 0.4equiv), and ammonium acetate (222.9 g, 4 equiv) were suspended in amixture of THF (720 mL) and MeOH (720 mL) in a 5 L round-bottomed flaskfitted with a magnetic stir bar and nitrogen inlet. Acetic acid (84 mL,2 equiv) was added and the mixture was heated in a 45° C. oil bath(solids dissolved upon heating). After 12 hours, LC/MS analysisindicated complete consumption of aldehyde starting material andformation of desired product (LC/MS m/z (M+H)⁺ 452.1). The MeOH/THF wereremoved by rotary evaporation. The residue was dissolved in 10% MeOH inCH₂Cl₂ (ca. 1.5 L) and the mixture was shaken vigorously with aqueouspotassium carbonate (ca. 210 g potassium carbonate in ca. 1.5 L water,pH of aqueous=8-9). The layers were separated, and the aqueous layer wasextracted with 10% MeOH in CH₂Cl₂ (2×100 mL). The combined organiclayers were concentrated to give a brown oily solid. The crude productwas suspended in 10% MeOH in EtOAc (ca. 1 L). Anhydrous Na₂SO₄ (ca. 60g) and silica gel (ca. 100 g) were added and the slurry was heatedgently with a heat gun to dissolve the crude product. The slurry wastransferred to a 2 L fritted glass filter funnel containing silica gel(ca. 100 g, pre-equilibrated with 10% MeOH in EtOAc), and the productwas eluted through the silica gel plug with 10% MeOH in EtOAc (ca. 6 L)and 1% Et₃N, 10% MeOH in EtOAc (ca. 10 L). (Note: Incomplete dissolutionof the product and/or precipitation of product in the presence of silicagel complicated the filtration). The solvents were removed by rotaryevaporation. The residue was triturated with MeCN (1×300 mL) and dried(rotary evaporation followed by high vacuum) to provide1-[4-(4-chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanoneas an off-white solid (190 g, 58%, LC/MS purity >98%).

Step 7:2-(3-1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanonehydrochloride salt

A 2 L flask with a magnetic stirrer was charged with the product of step6 (5.1 g, 11.28 mmol) and EtOAc (900 mL). The resulting suspension washeated to form a clear solution and cooled to room temperature undermoderate stirring. HCl in Et₂O (2M, 6.2 mL, 12.42 mmol) was addeddropwise to the resulting solution at room temperature over 5 min. Afterthe addition, the resultant suspension was stirred at room temperaturefor another 1 h. The solid was collected by filtration and washed withEt₂O (150 mL×2) and dried in vacuo to afford 5.4 g of an off-whitepowder. A 250 mL flask with a magnetic stirrer was charged with thepowder obtained above (5.4 g), acetone (100 mL) and deionized water (16mL). The resulting suspension was heated to form a clear solution andstirred to cool. When the solution became cloudy (crystal seedsappeared), acetone (540 mL) was added slowly to the suspension over 20min. The resulting suspension was heated to 50° C. and stirred for 2 h.Filtration while hot, washing with hot acetone (50 mL×2) and drying invacuo gave 3.3 g (60%) of the product as an off-white solid: m. p.164-165° C. The crystals appear as prisms under a polarizing microscope.

Example 79

Step 1: Synthesis of1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile

To a 2000 ml flask was charged with1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone(40 g, 78.1 mmol), dppf (3.86 g, 6.96 mmol), Zn(CN)₂ (9.6 g, 81.6 mmol),DMF (360 ml) and H₂O (20 ml). The resulting suspension was degassedusing N₂ for 5 min, followed by addition of Pd₂(dba)₃ (4.24 g, 4.64mmol). The reaction mixture was heated under N₂ at 90° C. for 2 h(monitor by TLC and LC-MS). After cooling to room temperature, dilutedwith EtOAc (1500 ml), filtered to remove the precipitate and washed withH₂O (1000×2 ml), saturated EDTA.4Na (800 ml×2), brine and dried overNa₂SO₄. After evaporation of the solvent, ether (150 mL) was added andstirred for 2 h. The resulting solid was filtered to give the desiredproduct 30 g (93%) as light yellow powder. Recrystallization fromrefluxing CH₃CN (160 mL) afforded 26 g (80%) light yellow crystals: mp183-185° C.; R_(t)=2.38 min; MS (ES) M+H expect 411.1. found 411.1.

Step 2: Synthesis of1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)-2-(3-(4,5-dihydro-1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)ethanone

A 250 mL flask was charged with1-(2-(4-chloro-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile(15.3 g, 37.2 mmol), EtOH (40 mL, ˜1 M). Under ice-bath and stirring,AcOH (6.75 mL, 112 mmol) was added, followed by ethylenediamine (25 mL,372 mmol). The resulting mixture was heated at 120° C. (bath) under N₂(observed mixture starting refluxing) for 1.5 h. TLC and LC-MS indicatedthe disappearance of starting material and formation of imidazoline.After cooling to room temperature, mixture was diluted with DCM (700 mL)and washed with H₂O (350 mL). The H₂O layer was back extracted with DCM(150 mL), and the combined organic layer was washed brine (350 mL) anddried over MgSO₄. After evaporating of the solvent under reducedpressure, the residue was suspended in hot EtOAc (80 mL). After coolingto room temperature, the solid was collected by filtration and washedwith EtOAc (30 mL) to afford the title compound as white powders (16 g,95%) which was used directly for next step: mp 133-135° C.; R_(t)=1.369min. MS (ES) M+H expect 454.2. found 454.4.

Step 3:2-(3-1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanone

The above imidazoline (12.3 g, 27.1 mmol) in a 500 mL flask was chargedwith anhydrous DMSO (108 mL, ˜0.25 M). DMP (17.2 g, 40.6 mmol) was addedin portions under stirring. The resulting mixture was stirred at 45° C.under N₂ for 2 h (monitor by TLC and LC-MS). After cooling to roomtemperature, quenching the reaction with sat Na₂S₂O₃ (100 mL)(ice-bath), followed 3 N NaOH (100 mL) (pH 12 to 13) and H₂O (300 mL)and extracted with DCM (600 mL+300 mL). The combined organic layer waswashed with sat NaHCO₃ (300 mL), brine (300 mL) and dried (MgSO₄, 120g). After evaporation of the organic solvent, the residue yellow solid(˜11 g) was dissolved in hot CH₃CN (20 mL). After cooling to roomtemperature, the resulting solid was collected by filtration to afford6.7 g (55%) of title compound as light tan crystals: mp 149-152° C.;R_(t)=1.309 min. MS (ES) M+H expect 452.2. found 452.4. Mother liquorwas concentrated and afforded another 0.6 g (total isolated yield 60%).

Example 80

This example illustrates the evaluation of the biological activityassociated with compounds of interest (candidate compounds) of theinvention.

Materials and Methods

A. Cells

1. CCR1 Expressing Cells

a) THP-1 Cells

THP-1 cells were obtained from ATCC (TIB-202) and cultured as asuspension in RPMI-1640 medium supplemented with 2 mM L-glutamine, 1.5g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1 mM sodiumpyruvate, 0.05% 2-mercaptoethanol and 10% PBS. Cells were grown under 5%CO₂/95% air, 100% humidity at 37° C. and subcultured twice weekly at 1:5(cells were cultured at a density range of 2×10⁵ to 2×10⁶ cells/mL) andharvested at 1×10⁶ cells/mL. THP-1 cells express CCR1 and can be used inCCR1 binding and functional assays.

b) Isolated Human Monocytes

Monocytes were isolated from human buffy coats using the Miltenyi beadisolation system (Miltenyi, Auburn, Calif.). Briefly, following a Ficollgradient separation to isolate peripheral blood mononuclear cells, cellswere washed with PBS and the red blood cells lysed using standardprocedures. Remaining cells were labeled with anti-CD14 antibodiescoupled to magnetic beads (Miltenyi Biotech, Auburn, Calif.). Labeledcells were passed through AutoMACS (Miltenyi, Auburn, Calif.) andpositive fraction collected. Monocytes express CCR1 and can be used inCCR1 binding and functional assays.

B. Assays

1. Inhibition of CCR1 Ligand Binding

CCR1 expressing cells were centrifuged and resuspended in assay buffer(20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl₂, 5 mM MgCl₂, and with 0.2%bovine serum albumin) to a concentration of 5×10⁶ cells/mL for THP-1cells and 5×10⁵ for monocytes. Binding assays were set up as follows.0.1 mL of cells (5×10⁵ THP-1 cells/well or 5×10⁴ monocytes) was added tothe assay plates containing the compounds, giving a final concentrationof ˜2-10 μM each compound for screening (or part of a dose response forcompound IC₅₀ determinations). Then 0.1 mL of ¹²⁵I labeled MIP-1α(obtained from Perkin Elmer Life Sciences, Boston, Mass.) or 0.1 mL of¹²⁵I labeled CCL15/leukotactin (obtained as a custom radiolabeling byPerkin Elmer Life Sciences, Boston, Mass.) diluted in assay buffer to afinal concentration of ˜50 pM, yielding ˜30,000 cpm per well, was added(using ¹²⁵I labeled MIP-1α with THP-1 cells and ¹²⁵I labeledCCL15/leukotactin with monocytes), the plates sealed and incubated forapproximately 3 hours at 4° C. on a shaker platform. Reactions wereaspirated onto GF/B glass filters pre-soaked in 0.3% polyethyleneimine(PEI) solution, on a vacuum cell harvester (Packard Instruments;Meriden, Conn.). Scintillation fluid (40 μl; Microscint 20, PackardInstruments) was added to each well, the plates were sealed andradioactivity measured in a Topcount scintillation counter (PackardInstruments). Control wells containing either diluent only (for totalcounts) or excess MIP-1α or MIP-1β (1 μg/mL, for non-specific binding)were used to calculate the percent of total inhibition for compound. Thecomputer program Prism from GraphPad, Inc. (San Diego, Calif.) was usedto calculate IC₅₀ values. IC₅₀ values are those concentrations requiredto reduce the binding of labeled MIP-1α to the receptor by 50%. (Forfurther descriptions of ligand binding and other functional assays, seeDairaghi, et al., J. Biol. Chem. 274:21569-21574 (1999), Penfold, etal., Proc. Natl. Acad. Sci. USA. 96:9839-9844 (1999), and Dairaghi, etal., J. Biol. Chem. 272:28206-28209 (1997)).

2. Calcium Mobilization

To detect the release of intracellular stores of calcium, cells (THP-1or monocytes) were incubated with 3 μM of INDO-1AM dye (MolecularProbes; Eugene, Oreg.) in cell media for 45 minutes at room temperatureand washed with phosphate buffered saline (PBS). After INDO-1AM loading,the cells were resuspended in flux buffer (Hank's balanced salt solution(HESS) and 1% FBS). Calcium mobilization was measured using a PhotonTechnology International spectrophotometer (Photon TechnologyInternational; New Jersey) with excitation at 350 nm and dualsimultaneous recording of fluorescence emission at 400 nm and 490 nm.Relative intracellular calcium levels were expressed as the 400 nm/490nm emission ratio. Experiments were performed at 37° C. with constantmixing in cuvettes each containing 10⁶ cells in 2 mL of flux buffer. Thechemokine ligands may be used over a range from 1 to 100 nM. Theemission ratio was plotted over time (typically 2-3 minutes). Candidateligand blocking compounds (up to 10 μM) were added at 10 seconds,followed by chemokines at 60 seconds (i.e., MIP-1α; R&D Systems;Minneapolis, Minn.) and control chemokine (i.e., SDF-1α; R&D Systems;Minneapolis, Minn.) at 150 seconds.

3. Chemotaxis Assays

Chemotaxis assays were performed using 5

m pore polycarbonate, polyvinylpyrrolidone-coated filters in 96-wellchemotaxis chambers (Neuroprobe; Gaithersburg, Md.) using chemotaxisbuffer (Hank's balanced salt solution (HESS) and 1% FBS). CCR1 chemokineligands (i.e., MIP-1α, CCL15/Leukotactin; R&D Systems; Minneapolis,Minn.) are use to evaluate compound mediated inhibition of CCR1 mediatedmigration. Other chemokines (i.e., SDF-1α; R&D Systems; Minneapolis,Minn.) are used as specificity controls. The lower chamber was loadedwith 29 μl of chemokine (i.e., 0.1 nM CCL15/Leukotactin) and varyingamounts of compound; the top chamber contained 100,000 THP-1 or monocytecells in 20 μl. The chambers were incubated 1-2 hours at 37° C., and thenumber of cells in the lower chamber quantified either by direct cellcounts in five high powered fields per well or by the CyQuant assay(Molecular Probes), a fluorescent dye method that measures nucleic acidcontent and microscopic observation.

C. Identification of Inhibitors of CCR1

1. Assay

To evaluate small organic molecules that prevent the receptor CCR1 frombinding ligand, an assay was employed that detected radioactive ligand(i.e., MIP-1α or CCL15/Leukotactin) binding to cells expressing CCR1 onthe cell surface (for example, THP-1 cells or isolated human monocytes).For compounds that inhibited binding, whether competitive or not, fewerradioactive counts are observed when compared to uninhibited controls.

THP-1 cells and monocytes lack other chemokine receptors that bind thesame set of chemokine ligands as CCR1 (i.e., MIP-1α, MPIF-1,Leukotactin, etc.). Equal numbers of cells were added to each well inthe plate. The cells were then incubated with radiolabeled MIP-1α.Unbound ligand was removed by washing the cells, and bound ligand wasdetermined by quantifying radioactive counts. Cells that were incubatedwithout any organic compound gave total counts; non-specific binding wasdetermined by incubating the cells with unlabeled ligand and labeledligand. Percent inhibition was determined by the equation:% inhibition=(1−[(sample cpm)−(nonspecific cpm)]/[(totalcpm)−(nonspecific cpm)])×100.

2. Dose Response Curves

To ascertain a candidate compound's affinity for CCR1 as well as confirmits ability to inhibit ligand binding, inhibitory activity was titeredover a 1×10⁻¹⁰ to 1×10⁻⁴ M range of compound concentrations. In theassay, the amount of compound was varied; while cell number and ligandconcentration were held constant.

3. CCR1 Functional Assays

CCR1 is a seven transmembrane, G-protein linked receptor. A hallmark ofsignaling cascades induced by the ligation of some such receptors is thepulse-like release of calcium ions from intracellular stores. Calciummobilization assays were performed to determine if the candidate CCR1inhibitory compounds were able to also block aspects of CCR1 signaling.Candidate compounds able to inhibit ligand binding and signaling with anenhanced specificity over other chemokine and non-chemokine receptorswere desired.

Calcium ion release in response to CCR1 chemokine ligands (i.e., MIP-1α,MPIF-1, Leukotactin, etc.) was measured using the calcium indicatorINDO-1. THP-1 cells or monocytes were loaded with INDO-1/AM and assayedfor calcium release in response to CCR1 chemokine ligand (i.e., MIP-1α)addition. To control for specificity, non-CCR1 ligands, specificallybradykinin, was added, which also signals via a seven transmembranereceptor. Without compound, a pulse of fluorescent signal will be seenupon MIP-1α addition. If a compound specifically inhibits CCR1-MIP-1αsignaling, then little or no signal pulse will be seen upon MIP-1αaddition, but a pulse will be observed upon bradykinin addition.However, if a compound non-specifically inhibits signaling, then nopulse will be seen upon both MIP-1α and bradykinin addition.

One of the primary functions of chemokines is their ability to mediatethe migration of chemokine receptor-expressing cells, such as whiteblood cells. To confirm that a candidate compound inhibited not onlyCCR1 specific binding and signaling (at least as determined by calciummobilization assays), but also CCR1 mediated migration, a chemotaxisassay was employed. THP-1 myelomonocytic leukemia cells, which resemblemonocytes, as wells as freshly isolated monocytes, were used as targetsfor chemoattraction by CCR1 chemokine ligands (i.e., MIP-1α,CCL15/leukotactin). Cells were place in the top compartment of amicrowell migration chamber, while MIP-1α (or other potent CCR1chemokine ligand) and increasing concentrations of a candidate compoundwas loaded in the lower chamber. In the absence of inhibitor, cells willmigrate to the lower chamber in response to the chemokine agonist; if acompound inhibited CCR1 function, then the majority of cells will remainin the upper chamber. To ascertain a candidate compound's affinity forCCR1 as well as to confirm its ability to inhibit CCR1 mediated cellmigration, inhibitory activity was titered over a 1×10⁻¹⁰ to 1×10⁻⁴ Mrange of compound concentrations in this chemotaxis assay. In thisassay, the amount of compound was varied; while cell number andchemokine agonist concentrations were held constant. After thechemotaxis chambers were incubated 1-2 hours at 37° C., the respondingcells in the lower chamber were quantified by labeling with the CyQuantassay (Molecular Probes), a fluorescent dye method that measures nucleicacid content, and by measuring with a Spectrafluor Plus (Tecan). Thecomputer program Prism from GraphPad, Inc. (San Diego, Calif.) was usedto calculate IC₅₀ values. IC₅₀ values are those compound concentrationsrequired to inhibit the number of cells responding to a CCR1 agonist by50%.

4. In Vivo Efficacy

a) Rabbit Model of Destructive Joint Inflammation

To study the effects of candidate compounds on inhibiting theinflammatory response of rabbits to an intra-articular injection of thebacterial membrane component lipopolysaccharide (LPS), a rabbit model ofdestructive joint inflammation is used. This study design mimics thedestructive joint inflammation seen in arthritis. Intra-articularinjection of LPS causes an acute inflammatory response characterized bythe release of cytokines and chemokines, many of which have beenidentified in rheumatoid arthritic joints. Marked increases inleukocytes occur in synovial fluid and in synovium in response toelevation of these chemotactic mediators. Selective antagonists ofchemokine receptors have shown efficacy in this model (see Podolin, etal., J. Immunol. 169(11):6435-6444 (2002)).

A rabbit LPS study is conducted essentially as described in Podolin, etal. ibid., female New Zealand rabbits (approximately 2 kilograms) aretreated intra-articularly in one knee with LPS (10 ng) together witheither vehicle only (phosphate buffered saline with 1% DMSO) or withaddition of CCX-105 (dose 1=50 μM or dose 2=100 μM) in a total volume of1.0 mL. Sixteen hours after the LPS injection, knees are lavaged andcells counts are performed. Beneficial effects of treatment weredetermined by histopathologic evaluation of synovial inflammation.Inflammation scores are used for the histopathologic evaluation:1—minimal, 2—mild, 3—moderate, 4—moderate-marked.

b) Evaluation of a Candidate Compound in a Rat Model of Collagen InducedArthritis

A 17 day developing type II collagen arthritis study is conducted toevaluate the effects of a candidate compound on arthritis inducedclinical ankle swelling. Rat collagen arthritis is an experimental modelof polyarthritis that has been widely used for preclinical testing ofnumerous anti-arthritic agents (see Trentham, et al., J. Exp. Med.146(3):857-868 (1977), Bendele, et al., Toxicologic Pathol. 27:134-142(1999), Bendele, et al., Arthritis Rheum. 42:498-506 (1999)). Thehallmarks of this model are reliable onset and progression of robust,easily measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation.

Female Lewis rats (approximately 0.2 kilograms) are anesthetized withisoflurane and injected with Freund's Incomplete Adjuvant containing 2mg/mL bovine type II collagen at the base of the tail and two sites onthe back on days 0 and 6 of this 17 day study. A candidate compound isdosed daily in a sub-cutaneous manner from day 0 till day 17 at aefficacious dose. Caliper measurements of the ankle joint diameter weretaken, and reducing joint swelling is taken as a measure of efficacy.

In the table below, structures and activity are provided forrepresentative compounds described herein. Activity is provided asfollows for either the chemotaxis assay or binding assay as describedabove: +, IC₅₀>12.5 uM; ++, 2500 nM<IC₅₀<12.5 uM; +++, 1000 nM<IC₅₀<2500nM; and ++++, IC₅₀<1000 nM.

TABLE 2 Structure

1. A method of treating rheumatoid arthritis comprising administering toa subject in need thereof a therapeutically effective amount of acompound having a formula selected from the group consisting of:

or a pharmaceutically acceptable salt, hydrate or N-oxide thereof,wherein the subscript m is an integer of from 0 to 2; each R¹ isindependently selected from the group consisting of —CO₂H and C₁₋₄alkyl, optionally substituted with —OH, —OR^(m), —S(O)₂R^(m), —CO₂H or—CO₂R^(m) wherein R^(m) is an unsubstituted C₁₋₆ alkyl; R^(2a), R^(2c)and R^(2d) are each independently selected from the group consisting ofhydrogen, halogen, cyano, oxazolyl, —NO₂, —CO₂R^(c), —CONR^(c)R^(d),—C(O)R^(c), —S(O)₂R^(e), —R^(e), —C(NOR^(c))R^(d), —OR^(c), —SR^(c),—NR^(d)C(O)R^(c), —X²OR^(c), —O—X²OR^(c), —X²NR^(c)R^(d),—NR^(c)S(O)₂R^(e), —OC(O)R^(c), and —NR^(e)R^(d); wherein within each ofR^(2a), R^(2c) and R^(2d), X² is C₁₋₄ alkylene and each R^(c) and R^(d)is independently selected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, andC₃₋₆ cycloalkyl and each R^(e) is independently selected from the groupconsisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, and C₃₋₆ cycloalkyl; each ofring vertices a, b, c and d in formulae Ia and Ib is independentlyselected from N and CH, and one of said ring vertices is N; and R^(3a)is selected from the group consisting of hydrogen, halogen,—NR^(f)R^(g), —R^(h), —S(O)₂R^(h), and —Y, wherein Y is selected fromthe group consisting of homopiperidinyl, morpholinyl, thiomorpholinyl,pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl,piperazinyl, phenyl, thienyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl,isothiazolyl, triazolyl, tetrazolyl and oxadiazolyl, optionallysubstituted with from one to three substitutents selected from the groupconsisting of halogen, —OR^(f), —NR^(f)R^(g), —R^(h), —CN, —NO₂,—CO₂R^(f), —CONR^(f)R^(g), and —C(O)R^(f), wherein each R^(f) and R^(g)is independently selected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, andC₃₋₆ cycloalkyl, and each R^(h) is independently selected from the groupconsisting of C₁₋₈ alkyl, C₁₋₈ haloalkyl, and C₃₋₆ cycloalkyl.
 2. Amethod in accordance with claim 1, wherein said administering is oral,parenteral, rectal, transdermal, sublingual, nasal or topical.
 3. Amethod in accordance with claim 1, wherein said compound is administeredin combination with an anti-inflammatory agent, analgesic agent, ananti-proliferative agent, a metabolic inhibitor, a leukocyte migrationinhibitor or an immuno-modulator.
 4. A method of treating rheumatoidarthritis comprising administering to a subject in need thereof atherapeutically effective amount of a compound selected from the groupconsisting of:1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyridin-1-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyridin-2-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4-b]pyridin-2-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone;2-(3-Amino-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[4,3-c]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-2-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-2-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-fluoro-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-2-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyridin-1-yl-2-oxide)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-morpholin-4-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-1-yl-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-c]pyridin-1-yl-6-oxide-ethanone;2-(3-Azidomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone;(1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridin-3-yl)-methanesulfonicacid;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-2-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(5-methyl-[1,2,4]oxadiazol-3-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methanesulfonyl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;2-(3-Aminomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-ethanone;1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;and1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone;or their pharmaceutically acceptable salts and N-oxides thereof.
 5. Amethod of treating rheumatoid arthritis comprising administering to asubject in need thereof a therapeutically effective amount of a compoundselected from the group consisting of:

or a pharmaceutically acceptable salt or N-oxide thereof.